HIV-1 group O antigens and uses thereof

ABSTRACT

The claimed invention relates to an HIV-1 group O envelope antigen comprising SEQ ID NO: 100, and the use of said antigen as a reagent in the diagnosis of HIV-1 group O infection, and a kit therefore.

The current invention relates to new HIV-1 group O antigens, nucleicacids encoding them, and the use of said antigens and/or nucleic acidsas reagents in the diagnosis and prophylaxes of AIDS. It also relates tonew HIV-1 group O strains comprising these antigens.

The human immunodeficiency virus (HIV) is the responsible agent for theacquired immunodeficiency syndrome (AIDS) in humans. AIDS is usuallyassociated with two distinct types of HIV: HIV-1 and HIV-2, initiallydescribed by Gallo et al. (1984) and Barré-Sinoussi et al. (1983) on theone hand, and Clavel et al. (1986) on the other hand. Although bothtypes, HIV-1 and HIV-2, cause a dysfunction of the immune system andinduce similar clinical symptoms in infected persons, they aregenetically distinct (Clavel et al. 1986) Epidemiological studies haveshown that the prevalence of HIV-2 infection is confined mainly to WestAfrica, whereas HIV-1 infection is a world wide problem. Numerous HIV-1isolates have been obtained and sequenced from diverse geographicallocations. At present, at least ten distinct subgroups or clades (A toJ) of HIV-1 have been described, equidistantly related in phylogeneticanalysis of the env-and/or gag-gene (Kostrikis et al. 1995; Louwagie etal. 1993; Myers et al 1995).

More recently, HIV-1 group O (for “Outlier”) strains have been describedas divergent viruses, belonging to an independent cluster (Charneau etal. 1994; Gürtler et al. 1994; Myers et al 1995; Sharp et al. 1994;Vanden Haesevelde et al. 1996), when compared to the vast majority ofworldwide HIV-1 strains classified as group M (for “Major”). Althoughthese two groups of viruses share the same genomic structure, theelevated level of divergence between them supports the hypothesis ofindependent origins.

Most of the currently described group O strains have been characterizedfrom Cameroonian patients or from patients who have travelled inCameroon (De Leys et al. 1990; Gürtler et al. 1994; Loussert-Ajaka etal. 1995; Vanden Haesevelde et al. 1996). Group O infection is notrestricted to Cameroon and its neighbouring countries, but it has alsobeen documented in West, East, and Southern Africa (Peeters et al. 1996;Peeters et al. submitted). In addition, cases of group O infection havebeen described in several European countries (France, Spain, Germany,Norway) and in the USA (Centres for Disease control and Prevention 1996;Charneau et al. 1994; Hampl et al 1995; Soriano et al. 1996).

Several hypotheses have been developed to explain the paradoxicalobservation that HIV-1 has been present in African countries for manydecades (probably about a century) and that it has only become apparentover the past 15 years. The answer should probably take in accountnumerous parameters such as demographic, sociologic, ethologic,ethnologic, and virologic parameters. In a mathematical model, May andAnderson (1990) suggest that initial chains of infection were found inisolated populations at low rates with some ‘sparks’ thrown in theneighbouring villages, and the exponential epidemic has started whenthere was a sufficient number of fire-boxes. To date, no differenceswere observed between HIV-1 group M and O pathogenic potential eventhough a limited number of patients infected by these latter strainshave been reported. However some of them have already died or reachedstage IV in the CDC classification (Charneau et al. 1994; Gürtler et al.1994; Loussert-Ajaka et al. 1995). It is possible that group Oepidemics, compared to group M, could be rampant at this time. In thenext years, it will therefore be extremely important to monitor theprevalence of these viruses, in Africa but also in the developedcountries, to detect them as early as possible and to prevent a new HIVepidemic.

HIV-1 group O strains present a public health challenge since they aredocumented to give incomplete and atypical HIV-1 Western blot profiles(Charneau et al. 1994; Gürtler et al. 1994). Some commercially availableELISA or rapid tests were unable to detect HIV-antibodies in HIV-1 groupO infected patients (Loussert-Ajaka et al 1994; Simon et al. 1994). Thedistribution of group O infections may be much more wide spread thancurrently thought, because of a lack of adequate detection techniques.Moreover, whereas HIV-1 group M strains have been extensively studiedand characterized as to their genetic variability, there is at presentno clear view on the genetic diversity of strains belonging to HIV-1group O.

At present, sequence information on the complete genome is onlyavailable for the prototype isolates of HIV-1 group O, namely ANT70(Vanden Haesevelde et al. 1994), MVP-5180 (Gürtler et al. 1994), and VAU(Charneau et al. 1994). Some additional HIV-1 group O strains have beensequenced in the gag and env regions (for example WO 96/27013, WO96/12809, EP 0727483).

HIV-viruses show a high degree of genetic variability. In the case ofHIV-1 viruses it is more or less accepted that at least one nucleotidechange occurs during one replication cycle. Certain regions of thegenome, for example those encoding structurally or enzymaticallyimportant proteins, may be rather conserved, but other regions,especially the env-region, may be subject of very high geneticvariability.

The envelope proteins of HIV are the viral proteins most accessible toimmune attack, and much attention has been directed towards elucidatingtheir structure and function. The env gene encoding the envelopeproteins consists of hypervariable sequences (V-regions) alternated bymore constant regions (C-regions) (Starcich et al, 1986; Willey et al,1986). The envelope protein is first synthesized as a heavilyglycosylated precursor protein (gp160), which is later cleaved by anon-viral protease to generate a transmembrane protein, also referred toas gp41, and an outer surface protein often referred to as gp120. Oneparticular region of the gp120 glycoprotein derived from the HIV-1 virustype has been studied extensively, namely the third hypervariable domain(V3) also known as the principal neutralizing determinant (PND)(Javaherian et al., 1989). The V3 domain of HIV-1 contains a loopstructure of 35 amino acids (V3-loop) which is formed by acysteine-cysteine disulfide bridge (Leonard et al. 1990). The gp41protein contains an immunodominant domain (ID) as found in allretroviruses. For HIV-viruses, this domain has been divided in twodistinct regions, corresponding to an immunosuppressive peptide (ISU) ofabout 17 aa, and a cysteine loop being the principal immunodominantdomain (PU)). The delineation of these respective regions in the gp41protein is demonstrated in FIG. 1.

The genetic variability of HIV-viruses considerably complicates bothdiagnosis and prevention of HIV-infection. Sera from patients infectedwith unknown types of HIV-virus, may contain antibodies which are notdetected by the current assay methods, which are based on (poly)peptidesequences of known viral strains. The detection of virus or viralantigen in certain samples, like organs for transplantation, or bloodtransfusion samples, may be missed due to the presence of hithertounknown variant types. Variation may occur in those genomic regionswhich are considered to be important in future vaccines. Finally, it isnot known at present if different genoric types may influence the courseof the AIDS disease, i.e. its virulence and/or susceptibility fortherapeutics.

Therefore, there is a constant need for characterization and sequencingof new HIV-strains, and especially of new HIV-1 group O strains, whichuntil now have only scarcely been characterized. Information on thegenetic variability of this “Outlier” group may enable a more rationalapproach for optimization of diagnostic tests and for development ofvaccines. Especially the variability of certain regions in the genome,known to be important target regions for the immune response, or forcertain therapeutic drugs, is of utmost importance. New sequencing datamay require the revision of existing diagnostic assays, and/or thedevelopment of new assays. Depending on the situation, it may beimportant to obtain a general detection of all HIV-infected samples,with a low number of false positives and false negatives, or to be ableto differentiate different types of HIV-infection (such as HIV-1 groupM, HIV-1 group O, HIV-2).

It is the aim of the current invention to provide new nucleic acid andpeptide sequences originating from HIV-1 group O strains.

It is more specifically the aim of the current invention to providenucleic acid and peptide sequences corresponding to the env-region ofnew HIV-1 group O strains, more particularly corresponding to the gp160env-precursor protein region, and most particularly to the C2V3 regionand the gp41 region.

It is also an aim of the present invention to provide for new viralstrains belonging to HIV-1 group O.

It is moreover an aim of the present invention to provide for antigensderived from said new HIV-1 group O strains.

It is also an aim of the current invention to provide for nucleic acidsderived from said new HIV-1 group O strains.

It is also an aim of the present invention to provide antibodiesreacting specifically with the antigens from the new HIV-1 group Ostrains.

It is moreover an aim of the present invention to provide for probeshybridizing specifically with the nucleic acids of the new HIV-1 group Ostrains.

It is moreover an aim of the present invention to use said antigensand/or antibodies and/or probes in a test for detecting the presence ofHIV-infection and/or to differentiate different types of HIV-infection.

It is thus also an aim of the present invention to provide for assaysenabling the detection and/or differentiation of HIV-infections.

It is finally also an aim of the present invention to provide forvaccine compositions providing protection against AIDS.

The following definitions serve to illustrate the terms and expressionsused in the different embodiments of the present invention as set outbelow:

The term “polynucleic acid” corresponds to either double-stranded orsingle-stranded cDNA or genomic DNA or RNA, containing at least 10, 20,30, 40 or 50 contiguous nucleotides. Single stranded polynucleic acidsequences are always represented in the current invention from the 5′end to the 3′ end.

Polynucleic acids according to the invention may be prepared by anymethod known in the art for preparing polynucleic acids (e.g. thephosphodiester method for synthesizing oligonucleotides as described byAgarwal et al. (1972), the phosphotriester method of Hsiung et al.(1979), or the automated diethylphosphoroamidite method of Baeucage etal. (1981)). Alternatively, the polynucleic acids of the invention maybe isolated fragments of naturally occurring or cloned DNA, cDNA or RNA.

The term “oligonucleotide” refers to a single stranded nucleic acidcomprising two or more nucleotides, and less than 100 nucleotides. Theexact size of an oligonucleotide depends on the ultimate function or useof said oligonucleotide. For use as a probe or primer theoligonucleotides are preferably about 5-50 nucleotides long, morepreferably 10-30 nucleotides long.

The oligonucleotides according to the present invention can be formed bycloning of recombinant plasmids containing inserts including thecorresponding nucleotide sequences, if need be by cleaving the latterout from the cloned plasmids upon using the adequate nucleases andrecovering them, e.g. by fractionation according to molecular weight.The probes according to the present invention can also be synthesizedchemically, e.g. by automatic synthesis on commercial instruments soldby a variety of manufacturers.

The nucleotides as used in the present invention may be ribonucleotides,deoxyribonucleotides and modified nucleotides such as inosine ornucleotides containing modified groups which do not essentially altertheir hybridisation characteristics. Moreover, it is obvious to the manskilled in the art that any of the below-specified probes can be used assuch, or in their complementary form, or in their RNA form (wherein T isreplaced by U).

The oligonucleotides used as primers or probes may also comprise orconsist of nucleotide analogues such as phosphorothioates (Matsukcura etal., 1987), alkylphosphorothioiates (Miller et al., 1979) or peptidenucleic acids (Nielsen et al., 1991; Nielsen et al., 1993) or maycontain intercalating agents (Asseline et al., 1984).

As most other variations or modifications introduced into the originalDNA sequences of the invention, these variations will necessitateadaptions with respect to the conditions under which the oligonucleotideshould be used to obtain the required specificity and sensitivity.However the eventual results of the hybridisation or amplification willbe essentially the same as those obtained with the unmodifiedoligonucleotides.

The introduction of these modifications may be advantageous in order topositively influence characteristics such as hybridization kinetics,reversibility of the hybrid-formation, biological stability of theoligonucleotide molecules, immobilization to solid phase etc.

The term “probe” refers to single stranded sequence-specificoligonucleotides which have a sequence which is sufficientlycomplementary to hybridize to the target sequence to be detected.

Preferably said probes are 90%, 95% or more homologous to the exactcomplement of the target sequence to be detected. These target sequencesmay be genornic DNA, genomic RNA or messenger RNA, or amplified versionsthereof.

The term “hybridizes to” refers to preferably stringent hybridizationconditions, allowing hybridisation between sequences showing at least90%, 95% or more homology with each other.

The term “primer” refers to a single stranded DNA oligonucleotidesequence capable of acting as a point of initiation for synthesis of aprimer extension product which is complementary to the nucleic acidstrand to be copied. The length and the sequence of the primer must besuch that they allow to prime the synthesis of the extension products.Preferably the primer is about 5-50 nucleotides long. Specific lengthand sequence will depend on the complexity of the required DNA or RNAtargets, as well as on the conditions of primer use such as temperatureand ionic strength. The fact that amplification primers do not have tomatch exactly with the corresponding template sequence to warrant properamplification is amply documented in the literature (Kwok et al., 1990).

The amplification method used can be either polymerase chain reaction(PCR; Saiki et al., 1988), ligase chain reaction (LCR; Landgren et al.,1988; Wu & Wallace, 1989, Barany, 1991), nucleic acid sequence-basedamplification (NASBA; Guatelli et al., 1990, Compton, 1991),transcription-based amplification system (TAS; Kwoh et al., 1989),strand displacement amplification (SDA; Duck, 1990; Walker et al., 1992)or amplification by means of Qβ replicase (Lizardi et al., 1988; Lomeliet al., 1989) or any other suitable method to amplify nucleic acidmolecules.

The term “complementary” nucleic acids as used in the current inventionmeans that the nucleic acid sequences can form a perfect base paireddouble helix with each other.

The terms “polypeptide” and “peptide” are used interchangeablythroughout the specification and designate a linear series of aminoacids connected one to the other by peptide bonds between thealpha-amino and carboxy groups of adjacent amino acids. Polypeptides canbe of a variety of lengths, either in their natural (uncharged) forms orin a charged form (=salt form), and either free of modifications such asglycosylation, side chain oxidation, or phosphorylation or containingthese modifications. Preferably the peptides of the invention are lessthan 100 amino acids in length, more preferably less than 50, and evenless than 30 amino acids long. It is well understood in the art thatamino acid sequences contain acidic and basic groups, and that theparticular ionization state exhibited by the peptide is dependent on thepH of the surrounding medium when the protein is in solution, or that ofthe medium from which it was obtained if the protein is in solid form.Also included in the definition are proteins modified by additionalsubstituents attached to the amino acids side chains, such as glycosylunits, lipids, or inorganic ions such as phosphates, as well asmodifications relating to chemical conversions of the chains, such asoxidation of sulfhydryl groups. Thus, “polypeptide” or its equivalentterms is intended to include the appropriate amino acid sequencereferenced, subject to those of the foregoing modifications which do notdestroy its functionality.

The polypeptides of the invention, and particularly the fragments, canbe prepared by classical chemical synthesis.

The synthesis can be carried out in homogeneous solution or in solidphase.

For instance, the synthesis technique in homogeneous solution which canbe used is the one described by Houbenweyl in the book entitled “Methodeder organischen chemie” (Method of organic chemistry) edited by E.Wunsh, vol. 15-I et II. THIEME, Stuttgart 1974.

The polypeptides of the invention can also be prepared in solid phaseaccording to the methods described by Atherton and Shepard in their bookentitled “Solid phase peptide synthesis” (IRL Press, Oxford, 1989).

The polypeptides according to this invention can also be prepared bymeans of recombinant DNA techniques as described by Maniatis et al.,Molecular Cloning: A Laboratory Manual, New York, Cold Spring HarborLaboratory, 1982). In that case the polypeptides are obtained asexpression products of the nucleic acids encoding said polypeptides. Theexpression occurs in a suitable host cell (eukaryotic or prokaryotic)which has been transformed with a vector in which the nucleic acidencoding the polypeptide has been inserted (called “insert”). Thenucleic acid insert may have been obtained through classical genomiccloning techniques (screening of genomic libraries, shotgun cloning etc. . . ), or by amplification of the relevant part in the viral genome,using suitable primer pairs and, for example, the polymerase chainreaction, or by DNA synthesis.

The word “antigen” refers to a molecule which provokes an immuneresponse (also called “immunogen”), or which can be recognized by theimmune system (also called “antigen sensu strictu”). The immune responseor the immune recognition reaction can be of the cellular or humoraltype. The antigens of the current invention are all polypeptides orpeptides, and therefore, the words “antigen” and “(poly)peptide” may beused interchangeably throughout the current invention.

The term “antigenic determinant” or “epitope” refers to that portion ofan antigenic molecule that is specifically bound by an antibodycombining site. Epitopes may be determined by any of the techniquesknown in the art or may be predicted by a variety of computer predictionmodels known from the art.

The terms “homologous” and “homology” are used in the current inventionas synonyms for “identical” and “identity”; this means that amino acidsequences which are e.g. said to be 55% homologous, show 55% identicalamino acids in the same position upon alignment of the sequences. Thesame definition holds for homologous nucleic acid sequences, i.e.nucleic acid sequences which are e.g. said to be 55% homologous, show55% identical base pairs in the same position upon alignment of thesequences.

The aims of the present invention have been met by the followingembodiments.

The present invention provides for an antigen, derived from thegp160-env precursor protein of a new HIV-1 group O strain, andcharacterized by an amino acid sequence comprising at least one of thefollowing sequences:

VQQMKI (SEQ ID NO 53),

KIGPMSWYSMG (SEQ ID NO 54),

GLEKN (SEQ ID NO 55),

IQQMKI (SEQ ID NO 56),

KIGPLAWYSMG (SEQ ID NO 57),

MGLERN (SEQ ID NO 58),

QSVQEIKI (SEQ ID NO 59),

KIGPMAWYSIG (SEQ ID NO 60),

IGIGTT (SEQ ID NO 61),

VQEIQT (SEQ ID NO 62),

QTGPMAWYSIH (SEQ ID NO 63),

IHLRTP (SEQ ID NO 64),

IQEIKI (SEQ ID NO 65),

KIGPMSWYSMG (SEQ ID NO 66),

MGIGQE (SEQ ID NO 67),

SVQELRI (SEQ ID NO 68),

RIGPMAWYSMT (SEQ ID NO 69),

MTLERD (SEQ ID NO 70),

SVQEIPI (SEQ ID NO 136),

and/or at least one amino acid sequence chosen from the following groupof sequences

RNQQLLNLWGCKGRLIC (SEQ ID NO 71),

CKGRLICYTSVQWNM (SEQ ID NO 72),

LWGCKGRIVC (SEQ ID NO 73),

SLWGCKGKLIC (SEQ ID NO 74),

CKGKSIC (SEQ ID NO 75),

CKGKIVC (SEQ ID NO 76),

CRGRQVC (SEQ ID NO 77),

CKGRLICYTSVH (SEQ ID NO 79),

CKGNLIC (SEQ ID NO 80),

CKGKMIC (SEQ ID NO 81),

CKGRVVC (SEQ ID NO 82),

or a fragment of said antigen, said fragment consisting of at least 8,preferably 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 50 up to the maximum number of contiguous amino acids of theamino acid sequence of said antigen, and being characterized by the factthat it specifically reacts with antibodies raised against said antigen.

The term “derived from” signifies that the antigen contains a fragmentof the gp160 env precursor protein.

The expression “specifically reacts with” means that the antigenfragment is specifically recognized by antibodies raised against theantigen from which it is derived. Specificity of reaction may bepreferably demonstrated using monoclonal antibodies raised against theantigen of the invention. Specificity of polyclonal antibodies may beobtained after absorption of said antibodies with the correspondingantigens of other HIV-1 group O strains, in order to eliminatenon-specific antibodies (=cross reactive antibodies) present in thepolyclonal mixture. The expression “specifically react with” also meansthat sera taken from patients infected with the HIV-1 group O strainfrom which the antigen of the invention originates, show a preferentialreaction with the antigen or antigen fragment of the invention, ascompared to the reactivity with a corresponding antigen or antigenfragment of other HIV-1 group O strains (=control), under comparablereaction conditions. This preferential reaction may be measuredquantitatively (e.g. ELISA absorption values) and should result inreactivity values which are at least 20%, 30%, 40% and preferably 50%higher than the reactivity with the control antigen. In practice, thismeans that the selected fragments of the above-mentioned antigens willalways show at least one amino acid difference when compared in analignment with the sequence of corresponding antigens of other HIV-1group O isolates, such as ANT70, MVP5180, VAU or others.

The above-mentioned amino acid sequences SEQ ID NO 53 to 70 and 136originate from the central region in the V3 loop of the gp160-envprecursor protein of new HIV-1 group O strains, while the amino acidsequences represented by SEQ ID NO 71-77 and 79-82 originate from thegp41-principal immunodominant domain (PID) of the gp160-env precursorprotein of the same HIV-1 group O strains.

The current invention also provides for antigens consisting of any ofthe amino acid sequences represented by SEQ ID NO 53-70, 136, 71-77,79-82, or consisting of an amino acid sequence according to any of SEQID NO 53-70, 136, 71-77, 79-82, whereby said sequence is extended at itsN-terminal and/or C-terminal end with at least 1, 2, 3, 4, 5, 6, 7, 8,9, 10, up to 15 amino acids.

The invention further provides for an antigen as described above,characterized by an amino acid sequence comprising at least one of thefollowing amino acid sequences:

CERPGNNSIQQMKIGPLAWYSMGLERNKSSISRLAYC (SEQ ID NO 83),

CERPGNNSIQQMKIGPMAWYSMGLERNKSSISRLAYC (SEQ ID NO 84),

CERPGNQSVQEIKIGPMAWYSIGIGTTPANWSRIAYC (SEQ ID NO 85),

CERPGNQSVQEIKIGPMAWYSIGIGTTPTYNWSRIAYC (SEQ ID NO 86),

CVRPWNQTVQEIQTGPMAWYSIHLRTPLANLSRIAYC (SEQ ID NO 87),

CQRPGNLTIQEIKIGPMSWYSMGIGQEDHSKSRNAYC (SEQ ID NO 88),

CERPYYQSVQELRIGPMAWYSMTLERDRAGSDIRAAYC (SEQ ID NO 89),

CERPGNHTVQQMKIGPMSWYSMGLEKNNTSSRRAFC (SEQ ID NO 90),

CERTWNQSVQEIPIGPMAWYSMSVELDLNTTGSRSADC (SEQ ID NO 135),

and/or at least one amino acid sequence chosen from the following groupof sequences:

DQQLLNLWGCKGRIVCYTSVKWN (SEQ ID NO 91),

NQQLLNLWGCKGRLVCYTSVKWNK (SEQ ID NO 92),

NQQLLNLWGCKGRLVCYTSVKWNN (SEQ ID NO 138),

NQQRLNLWGCKGKMICYTSVPWN (SEQ ID NO 93),

NQQLLNLWGCKGKSICYTSVKWN (SEQ ID NO 94),

NQQLLNLWGCKGRLICYTSVQWN (SEQ ID NO 95),

NQQRLNLWGCKGKMICYTSVKWN (SEQ ID NO 96),

NQQLLNLWGCKGNLICYTSVKWN (SEQ ID NO 97),

NQQLLNLWGCRGRQVCYTSVIWN (SEQ ID NO 98),

SQQLLNLWGCKGRLICYTSVHWN (SEQ ID NO 99),

NQQLLNLWGCKGRIVCYTSVKWN (SEQ ID NO 100),

NQQLLNSWGCKGKIVCYTAVKWN (SEQ ID NO 101),

NQQLLSLWGCKGKLICYTSVKWN (SEQ ID NO 102),

NQQLLNLWGCKGRLVCYTSVQWN (SEQ ID NO 137),

or a fragment of said antigen, said fragment consisting of at least 8,preferably 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 50 up to the maximum number of contiguous amino acids of theamino acid sequence of said antigen, and being characterized by the factthat it specifically reacts with antibodies raised against said antigen.

The above-mentioned amino acid sequences SEQ ID NO 83 to 90 and 135represent the V3 loop region of the gp160-env precursor protein of newHIV-1 group O strains, while the amino acid sequences SEQ ID NO 91 to102, 137 and 138 originate from the gp41-immunodominant domain (ID) ofthe gp160-env precursor protein of the same HIV-1 group O strains.

The current invention also provides for antigens consisting of any ofthe amino acid sequences represented by SEQ ID NO 83-102, 135, 137 and138 or consisting of an amino acid sequence according to any of SEQ IDNO 83-102, 135, 137 and 138, whereby said sequence is extended at itsN-terminal and/or C-terminal end with at least 1, 2, 3, 4, 5, 6, 7, 8,9, 10, up to 15 amino acids.

The invention further provides for antigens as above-defined,characterized by an amino acid sequence comprising at least one of theamino acid sequences represented by SEQ ID NO 2, SEQ ID NO 4, SEQ ID NO6, SEQ ID NO 8, SEQ ID NO 10, SEQ ID NO 12, SEQ ID NO 14, SEQ ID NO 16,SEQ ID NO 18, SEQ ID NO 20, SEQ ID NO 22, SEQ ID NO 24, SEQ ID NO 26,SEQ ID NO 28, SEQ ID NO 30, SEQ ID NO 32, SEQ ID NO 34, SEQ ID NO 36,SEQ ID NO 38, SEQ ID NO 40 as shown in the alignment on FIG. 1, and/orat least one of the amino acid sequences represented by SEQ ID NO 42,SEQ ID NO 44, SEQ ID NO 46, SEQ ID NO 48, SEQ ID NO 50, or SEQ ID NO 52as shown in the alignment on FIG. 2, and/or the amino acid sequencerepresented by SEQ ID NO 134, or a fragment of said antigen, saidfragment consisting of at least 8, preferably 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up to the maximum number ofcontiguous amino acids of any of the sequences represented by SEQ ID NO2, SEQ ID NO 4, SEQ ID NO 6, SEQ ID NO 8, SEQ ID NO 10, SEQ ID NO 12,SEQ ID NO 14, SEQ ID NO 16, SEQ ID NO 18, SEQ ID NO 20, SEQ ID NO 22,SEQ ID NO 24, SEQ ID NO 26, SEQ ID NO 28, SEQ ID NO 30, SEQ ID NO 32,SEQ ID NO 34, SEQ ID NO 36, SEQ ID NO 38, SEQ ID NO 40, SEQ ID NO 42,SEQ ID NO 44, SEQ ID NO 46, SEQ ID NO 48, SEQ ID NO 50, SEQ ID NO 52, orSEQ ID NO 134, with said antigen fragment characterized by the fact thatit specifically reacts with antibodies raised against the antigen fromwhich it is derived.

Furthermore, the invention provides for an antigen as above-defined,characterized by an amino acid sequence consisting of at least one ofthe following sequences: SEQ ID NO 2, SEQ ID NO 4, SEQ ID NO 6, SEQ IDNO 8, SEQ ID NO 10, SEQ ID NO 12, SEQ ID NO 14, SEQ ID NO 16, SEQ ID NO18, SEQ ID NO 20, SEQ ID NO 22, SEQ ID NO 24, SEQ ID NO 26, SEQ ID NO28, SEQ ID NO 30, SEQ ID NO 32, SEQ ID NO 34, SEQ ID NO 36, SEQ ID NO38, SEQ ID NO 40, SEQ ID NO 42, SEQ ID NO 44, SEQ ID NO 46, SEQ ID NO48, SEQ ID NO 50, SEQ ID NO 52, or the amino acid sequence representedby SEQ ID NO 134 or a fragment of said antigen, said fragment consistingof at least 8, preferably 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,21, 22, 23, 24, 25, 50 up to the maximum number of contiguous aminoacids of any of the sequences represented by SEQ ID NO 2, SEQ ID NO 4,SEQ ID NO 6, SEQ ID NO 8, SEQ ID NO 10, SEQ ID NO 12, SEQ ID NO 14, SEQID NO 16, SEQ ID NO 18, SEQ ID NO 20, SEQ ID NO 22, SEQ ID NO 24, SEQ IDNO 26, SEQ ID NO 28, SEQ ID NO 30, SEQ ID NO 32, SEQ ID NO 34, SEQ ID NO36, SEQ ID NO 38, SEQ ID NO 40, SEQ ID NO 42, SEQ ID NO 44, SEQ ID NO46, SEQ ID NO 48, SEQ ID NO 50, SEQ ID NO 52, or SEQ ID NO 134, withsaid antigen fragment characterized by the fact that it specificallyreacts with antibodies raised against the antigen from which it isderived.

It is to be noted that all the above-mentioned amino acid sequencesoriginate from HIV-1 group O strains, which have until now never beendescribed. More particularly, as is shown further in the examplessection, the new amino acid sequences originate from the followingstrains:

The amino acid sequences represented by SEQ ID NO 2, 4, 42, 73, 59, 60,61, 73, 85, 86, 100 originate from the gp160 env precursor antigenisolated from a HIV-1 group O strain termed MP340, or a quasi-speciesthereof.

The amino acid sequences represented by SEQ ID NO 6, 8, 44, 56, 57, 58,82, 83, 84, 138 originate from the gp160 env precursor antigen isolatedfrom a HIV-1 group O strain termed FABA, or alternatively termed MP331,or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 10, 12, 46, 62, 63,64, 73, 87, 100 originate from the gp160 env precursor antigen isolatedfrom a HIV-1 group O strain termed MP450, or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 14, 16, 48, 65, 66,67, 76, 88, 101 originate from the gp160 env precursor antigen isolatedfrom a HIV-1 group O strain termed MP448, or a quasi species thereof.

The amino acid sequences represented by SEQ ID NO 18, 50, 53, 54, 55,73, 90, 91 originate from the gp160 env precursor antigen isolated froma HIV-1 group O strain termed 189, or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 40, 52, 68, 69, 70,71, 89, 95 originate from the gp160 env precursor antigen isolated froma HIV-1 group O strain termed MP539, or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 20 and 92 originatefrom the gp160 env precursor antigen isolated from a HIV-1 group Ostrain termed 320, or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 22, 80 and 97originate from the gp160 env precursor antigen isolated from a HIV-1group O strain termed BSD422, or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 24, 79 and 99originate from the gp160 env precursor antigen isolated from a HIV-1group O strain termed KGT008, or a quasi-species thereof.

The amino acid sequence represented by SEQ ID NO 26 originates from thegp160 env precursor antigen isolated from a HIV-1 group O strain termedMP575, or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 28, 72 and 95originate from the gp160 env precursor antigen isolated from a HIV-1group O strain termed BSD189, or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 30, 77 and 98originate from the gp160 env precursor antigen isolated from a HIV-1group O strain termed BSD649, or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 32, 81 and 96originate from the gp160 env precursor antigen isolated from a HIV-1group O strain termed BSD242, or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 34, 81 and 93originate from the gp160 env precursor antigen isolated from a HIV-1group O strain termed 533, or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 36, 75 and 94originate from the gp160 env precursor antigen isolated from a HIV-1group O strain termed 772P94, or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 38, 74 and 102originate from the gp160 env precursor antigen isolated from a HIV-1group O strain termed MP95B, or a quasi-species thereof.

The amino acid sequences represented by SEQ ID NO 134, 135, 136, and 137originate from the gp160 env precursor antigen isolated from a HIV-1group O strain termed MP645, or a quasi-species thereof.

It is noted that the amino acid sequence represented by SEQ ID NO 73 ischaracteristic for the gp41 immunodominant region of at least thefollowing new HIV-1 group O strains: MP340, MP450, and 189.

The current invention therefore specifically relates to env-derivedantigens comprising the characteristic sequence represented by SEQ ID NO73, as well as virus strains containing these antigens.

It is also noted that the amino acid sequence represented by SEQ ID NO81 is characteristic for the gp41 immunodominant region of at least thefollowing new HIV-1 group O strains: BSD242 and 533.

The current invention therefore specifically relates to env-derivedantigens comprising the characteristic sequence represented by SEQ ID NO81, as well as virus strains containing these antigens.

It is also noted that the amino acid sequence represented by SEQ ID NO95 is characteristic for the gp41 immunodominant region of at least thefollowing new HIV-1 group O strains: MP539 and BSD189.

The current invention therefore specifically relates to env-derivedantigens comprising the characteristic sequence represented by SEQ ID NO95, as well as virus strains containing these antigens.

The term “quasi-species” refers in general to the group of related butgenetically and possibly biologically different viruses (also called“variants”) that an infected individual harbors. The term “related”means that the “variants” all arise from a single infectious agent, inthis case from a single HIV-1 group O strain. It has been calculatedthat an HIV-infected patient carries about 10⁶ to 10⁸ geneticallydistinct HIV-variants, which are generated by the high error rate ofreverse transcriptase and the high turnover rate in vivo. In the contextof the current description the term “quasi-species” refers also to astrain isolated from the quasi-species “group” as above-defined.

The term “genetically different” means that the nucleic acid sequence ofthe genome of one strain shows at least one nucleotide difference withthe corresponding sequence of another strain belonging to the samequasi-species.

The term “biologically different” means that some strains of aquasi-species may have different biological characteristics compared tothe biological characteristics of other strains from the samequasi-species. These biological characteristics may encompass forexample the HIV-1 cell tropism, viral virulence, the capacity to inducesyncytia, etc.

Nucleic acid sequences originating from quasi-species differ from eachother but always show a high percentage of homology, most often ahomology of 90%, 95% or higher. The same holds for the sequence ofpolypeptides originating from quasi-species. Homology percentages on theprotein level usually exceed 95%, 96%, 97%, 98%, or even 99%. Thesepercentages of homology count for the comparison of sequence stretcheswhich are at least 100 nucleotides (about 33 amino acids), andpreferably 200, 300 or more nucleotides long (66, 100 or more aminoacids). It has to be understood that, when very short sequence stretchesare compared (e.g. stretches of about 30 nucleotides, or 10 amino acids)the homology ranges may be much lower, if these short sequence stretchescontain the mutual differences.

Examples of sequences originating from “quasi-species” are providedfurther in the examples section, where gp41- and C2V3-nucleotide andamino acid sequences of certain strains belonging to the same“quasi-species” are compared to each other. For example, for strainsMP340, FABA, MP450 and MP448 gp41-nucleic acid sequences have beendetermined on different samples, originating from the same patient, i.e.on serum samples and on peripheral blood monocyte (PBMC) samples. Table2 shows that, in these specific examples, homology percentages vary from95% to 100% between gp41-nucleic acid sequences determined on serumsamples as compared to PBMC-samples.

It is to be understood that the amino acid and nucleic acid sequences ofthe current invention also encompass those sequences which are notexplicitly recited, but which have been determined on “quasi-species” ofthe respective viral strains. As indicated above, these “variant”sequences show a homology range of at least 90%, preferably 95% with thesequences which are specifically recited in the current application.

The above-mentioned antigens are polypeptide or peptide molecules, whichare characterized by the above-mentioned amino acid sequences. It has tobe understood however, that these (poly)peptides may be modified by forexample glycosylation, side chain oxidation or phosphorylation asexplained above. A very particular type of side chain oxidation iscyclisation by bridge formation between the —SH groups of two cysteineresidues in the same (poly)peptide chain. The cyclic (poly)peptidesformed in this way by S—S bridging may be particularly suitable toexpose epitopes located in the loop structure. Epitopes presented inthis manner may be in a better shape to be recognized by the immunesystem, and more particularly by antibodies possibly present in theserum of HIV-infected persons.

A preferential embodiment of the current invention provides for any ofthe above-mentioned (poly)peptides in a cyclic form.

Cyclisation may occur between two cysteine residues which are present inthe above-cited amino acid sequences. For example, cyclic peptides witha loop structure of about 6 amino acids long may be formed with theamino acid sequences represented by e.g. SEQ ID NO 71 to 82, and 91 to102, and 137. Another example are the V3-loop peptides of about 35 aminoacids long, which may be formed by cyclisation of the cysteine residuesof the amino acid sequences represented by e.g. SEQ ID NO 83 to 90, and135.

On the other hand, cyclisation may also be induced in amino acidsequences which do not contain two cysteine residues naturally, butwhich have been extended with one or two cysteine residues at theirextremities, or at in internal position inside the amino acid chain. Thecurrent invention therefore also refers to (poly)peptides characterizedby any of the above-mentioned amino acid sequences, modified by additionof one or several cysteine residues, at the C-terminal and/or N-terminalextremity and/or inside the (poly)peptide chain.

Another particular type of modification includes the extension of theN-terminal and/or C-terminal end of the (poly)peptide antigen by linkersequences, said linker sequences comprising for example additional aminoacids or other molecules (such as for example biotin). The addition oflinker sequences to the polypeptide antigen may have several advantagessuch as:

a more efficient immobilisation on a solid substrate,

a more efficient presentation of the immunoreactive epitope(s) in the(poly)peptide,

linkage to other antigenic determinants . . . .

A preferential embodiment therefore includes antigens or antigenfragments comprising any of the above-mentioned amino acid sequences,extended with linker sequences.

It has to be understood that the above-mentioned (poly)peptide antigensof the invention may be prepared by different methods known in the art.They may be prepared by synthetic means as described above, or they maybe produced by recombinant DNA technology. In the latter case, they arethe result of the expression of the nucleic acids encoding said antigensor antigen fragments in an appropriate host cell.

The invention also relates to a recombinant vector for the expression ofany of the above-mentioned polypeptides, recombinant host cellsexpresssing these polypeptides, and processes for the recombinantexpression of these polypeptides; said tools for recombinant expressionare well known by anyone skilled in the art, and have been described inmore detail for example in WO96/13590.

The invention further provides for a (poly)nucleic acid encoding any ofthe above-mentioned (poly)peptide antigens.

More particularly, the current invention provides for a polynucleic acidcomprising a nucleotide sequence chosen from the group of

(I) a nucleotide sequence represented by SEQ ID NO 1, SEQ ID NO 3, SEQID NO 5, SEQ ID NO 7, SEQ ID NO 9, SEQ ID NO 11, SEQ ID NO 13, SEQ ID NO15, SEQ ID NO 17, SEQ ID NO 19, SEQ ID NO 21, SEQ ID NO 23, SEQ ID NO25, SEQ ID NO 27, SEQ ID NO 29, SEQ ID NO 31, SEQ ID NO 33, SEQ ID NO35, SEQ ID NO 37, SEQ ID NO 39, SEQ ID NO 41, SEQ ID NO 43, SEQ ID NO45, SEQ ID NO 47, SEQ ID NO 49, or SEQ ID NO 51, SEQ ID NO 106 or

(ii) a nucleotide sequence complementary to a sequence according to (I),or

(iii) a nucleotide sequence showing at least 95%, preferably 96%, 97%,98% and most preferably 99% homology to a sequence according to (I), or

(iv) a nucleotide sequence according to (I) whereby T is replaced by U,or

(v) a nucleotide sequence according to (I) whereby at least onenucleotide is substituted by a nucleotide analogue.

It is to be noted that, as will be shown further on in the examplessection, the above-mentioned polynucleic acids all originate from theenv-gene of new HIV-1 group O strains. The nucleic acid sequencesrepresented by SEQ ID NO 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25,27, 29, 31, 33, 35, 37 and 39 correspond to the region encoding thegp41-immunodominant domain in the env-gene, while the nucleic acidsequences represented by SEQ ID NO 41, 43, 45, 47, 49, and 51 correspondto the region encoding the C2V3 region in this same env-gene. Thenucleotide sequence represented by SEQ ID NO 106 is illustrated in FIG.8A, and comprises the full env-gene of a new HIV-1 group O strain,termed MP645, together with additional accompanying genes.

The nucleotide sequences mentioned above under item (iii) representvariant nucleic acid sequences which may be isolated e.g. from strainsbelonging to the same quasi-species.

More particularly, the invention provides for a polynucleic acidconsisting of a nucleotide sequence chosen from the group of

(I) a nucleotide sequence represented by SEQ ID NO 1, SEQ ID NO 3, SEQID NO 5, SEQ ID NO 7, SEQ ID NO 9, SEQ ID NO 11, SEQ ID NO 13, SEQ ID NO15, SEQ ID NO 17, SEQ ID NO 19, SEQ ID NO 21, SEQ ID NO 23, SEQ ID NO25, SEQ ID NO 27, SEQ ID NO 29, SEQ ID NO 31, SEQ ID NO 33, SEQ ID NO35, SEQ ID NO 37, SEQ ID NO 39, SEQ ID NO 41, SEQ ID NO 43, SEQ ID NO45, SEQ ID NO 47, SEQ ID NO 49, or SEQ ID NO 51, SEQ ID NO 106 or

(ii) a nucleotide sequence complementary to a sequence according to (I),or

(iii) a nucleotide sequence showing at least 95%, preferably 96%, 97%,98% and most preferably 99% homology to a sequence according to (I), or

(iv) a nucleotide sequence according to (I) whereby T is replaced by U,or

(v) a nucleotide sequence according to (I) whereby at least onenucleotide is substituted by a nucleotide analogue.

The invention further provides for a nucleic acid fragment consisting ofa sequence of at least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23,24, 25 up to 50 contiguous nucleotides of the sequence of a polynucleicacid as specified above, and characterized by the fact that itselectively hybridizes to the polynucleic acid from which it is derived.

The above-described nucleic acid fragment may be used as a specifichybridization probe for the detection of the nucleic acids of thecurrent invention.

The term “selectively hybridizing” means that the hybridization signalobtained after hybridization of the fragment with the nucleic acid fromwhich it is derived, is more intense than the hybridization signalobtained when the fragment is hybridized to the corresponding nucleicacid from another HIV-1 group O strain, under the same stringenthybridization and wash conditions. In practice this means that thenucleic acid fragment will show at least one mismatched nucleotide withthe sequence of the corresponding nucleic acid fragment of another HIV-1group O strain.

The term “stringent hybridization conditions” implies that thehybridization takes place at a temperature which is situatedapproximately between Tm and (Tm−10° C.), whereby Tm represents thecalculated melting temperature of the target nucleic. It is generallyknown that the stringency depends on the percentage mismatches(=non-matching nucleotides upon alignment) present in the hybridizingduplex. According to a simplified formula, the hybridization temperaturemay be calculated as follows: Tm−1.2 (% mismatch). A temperaturedecrease of 10° C. implies a maximum percentage of allowed mismatches of8.3%.

The invention further provides for a nucleic acid fragment consisting ofa sequence of at least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23,24, 25 up to 50 contiguous nucleotides of the sequence of a polynucleicacid as specified above, and characterized by the fact that itselectively amplifies the polynucleic acid from which it is derived.

The nucleic acid fragment as described above may be used as a specificamplification primer of the nucleic acids of the current invention.

The term “selective amplification” refers to the fact that said nucleicacid fragment may initiate a specific amplification reaction of thenucleic acids of the invention (e.g. a polymerase chain reaction) in thepresence of other nucleic acids, under appropriate amplificationconditions. It means that, under the appropriate amplificationconditions, only the nucleic acids of the invention will be amplified,and not the other nucleic acids possibly present.

Preferred embodiments of the invention comprise polynucleic acids orfragments thereof. as specified below.

A polynucleic acid comprising SEQ ID NO 1, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 1.

A polynucleic acid consisting of SEQ ID NO 1, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 1, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 1.

A polynucleic acid comprising SEQ ID NO 3, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 3.

A polynucleic acid consisting of SEQ ID NO 3, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 3, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 3.

A polynucleic acid comprising SEQ ID NO 5, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 5.

A polynucleic acid consisting of SEQ ID NO 5, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 5, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 5.

A polynucleic acid comprising SEQ ID NO 7, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 7.

A polynucleic acid consisting of SEQ ID NO 7, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 7, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 7.

A polynucleic acid comprising SEQ ID NO 9, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 9.

A polynucleic acid consisting of SEQ ID NO 9, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 9, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 9.

A polynucleic acid comprising SEQ ID NO 11, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 11.

A polynucleic acid consisting of SEQ ID NO 11, or a fragment consistingof at least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, up to50 contiguous nucleotides of SEQ ID NO 11, said fragment characterizedby the fact that it selectively hybridizes to the polynucleic acidconsisting of SEQ ID NO 11.

A polynucleic acid comprising SEQ ID NO 13, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 13.

A polynucleic acid consisting of SEQ ID NO 13, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, upto the maximum number of contiguous nucleotides of SEQ ID NO 13, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 13.

A polynucleic acid comprising SEQ ID NO 15, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 15.

A polynucleic acid consisting of SEQ ID NO 15, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 15, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 15.

A polynucleic acid comprising SEQ ID NO 17, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 17.

A polynucleic acid consisting of SEQ ID NO 17, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 17, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 17.

A polynucleic acid comprising SEQ ID NO 19, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 19.

A polynucleic acid consisting of SEQ ID NO 19, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 19, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 19.

A polynucleic acid comprising SEQ ID NO 21, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 21.

A polynucleic acid consisting of SEQ ID NO 21, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 21, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 21.

A polynucleic acid comprising SEQ ID NO 23, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 23.

A polynucleic acid consisting of SEQ ID NO 23, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 23, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 23.

A polynucleic acid comprising SEQ ID NO 25, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 25.

A polynucleic acid consisting of SEQ ID NO 25, or a fragment consistingof at least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, up to50 contiguous nucleotides of SEQ ID NO 25, said fragment characterizedby the fact that it selectively hybridizes to the polynucleic acidconsisting of SEQ ID NO 25.

A polynucleic acid comprising SEQ ID NO 27, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 27.

A polynucleic acid consisting of SEQ ID NO 27, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 27, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 27.

A polynucleic acid comprising SEQ ID NO 29, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 29.

A polynucleic acid consisting of SEQ ID NO 29, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 29, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 29.

A polynucleic acid comprising SEQ ID NO 31, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 31.

A polynucleic acid consisting of SEQ ID NO 31, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 31, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 31.

A polynucleic acid comprising SEQ ID NO 33, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 33.

A polynucleic acid consisting of SEQ ID NO 33, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 33, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 33.

A polynucleic acid comprising SEQ ID NO 35, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 35.

A polynucleic acid consisting of SEQ ID NO 35, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 35, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 35.

A polynucleic acid comprising SEQ ID NO 37, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 37.

A polynucleic acid consisting of SEQ ID NO 37, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 37, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 37.

A polynucleic acid comprising SEQ ID NO 39, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 39.

A polynucleic acid consisting of SEQ ID NO 39, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 39, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 39.

A polynucleic acid comprising SEQ ID NO 41, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 41.

A polynucleic acid consisting of SEQ ID NO 41, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 41, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 41.

A polynucleic acid comprising SEQ ID NO 43, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 43.

A polynucleic acid consisting of SEQ ID NO 43, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 43, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 43.

A polynucleic acid comprising SEQ ID NO 45, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 45.

A polynucleic acid consisting of SEQ ID NO 45, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 45, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 45.

A polynucleic acid comprising SEQ ID NO 47, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 47.

A polynucleic acid consisting of SEQ ID NO 47, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 47, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 47.

A polynucleic acid comprising SEQ ID NO 49, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 49.

A polynucleic acid consisting of SEQ ID NO 49, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 49, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 49.

A polynucleic acid comprising SEQ ID NO 51, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 51.

A polynucleic acid consisting of SEQ ID NO 51, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 51, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 51.

A polynucleic acid comprising SEQ ID NO 106, or a fragment consisting ofat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 up to 50contiguous nucleotides of said polynucleic acid, said fragmentcharacterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 106.

A polynucleic acid consisting of SEQ ID NO 106, or a fragment comprisingat least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up tothe maximum number of contiguous nucleotides of SEQ ID NO 106, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid consisting of SEQ ID NO 106.

The invention further provides for a virus strain belonging to HIV-1group O, comprising in its genome any of the above-mentioned nucleicacids.

More particularly, the invention provides for a virus strain belongingto HIV-1 group O, comprising in its genome the RNA equivalent of

one of the DNA sequences represented by SEQ ID NO 1, SEQ ID NO 3, SEQ IDNO 5, SEQ ID NO 7, SEQ ID NO 9, SEQ ID NO 11, SEQ ID NO 13, SEQ ID NO15, SEQ ID NO 17, SEQ ID NO 19, SEQ ID NO 21, SEQ ID NO 23, SEQ ID NO25, SEQ ID NO 27, SEQ ID NO 29, SEQ ID NO 31, SEQ ID NO 33, SEQ ID NO35, SEQ ID NO 37, SEQ ID NO 39, SEQ ID NO 106 and/or

one of the DNA sequences represented by SEQ ID NO 41, SEQ ID NO 43, SEQID NO 45, SEQ ID NO 47, SEQ ID NO 49, SEQ ID NO 51, and/or

a variant sequence of the above-mentioned DNA sequences, said variantsequence showing at least 95% homology with the entire length of one ofthe above-mentioned sequences.

More particularly, the invention relates to a strain of HIV-1 group O asdefined above, comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 5 and/or SEQ ID NO 43 or a variantsequence thereof, said variant sequence showing at least 95% homologywith SEQ ID NO 5 and/or SEQ ID NO 43. An HIV-1 group O strain of thistype, termed FABA (or synonymously MP331) has been deposited at theECACC on Jun. 13 1997, under accession No V97061301.

An example of a variant sequence of SEQ ID NO 5 is SEQ ID NO 7. Thelatter sequence was determined on a serum sample of a patient infectedby the strain FABA, while the former sequence was determined onperipheral blood mononuclear cells (PBMC's) taken from the same patient.The nucleic acids represented by SEQ ID NO 5 and SEQ ID NO 7 show 95%homology, and can be said to belong to strains from the samequasi-species.

The invention also relates to a strain of HIV-1 group O as definedabove, comprising in its genome the RNA equivalent of the DNA sequencerepresented by SEQ ID NO 9 and/or SEQ ID NO 45 or a variant sequence,said variant sequence showing at least 95% homology with SEQ ID NO 9and/or SEQ ID NO 45. An HIV-1 group O strain of this type, termed MP450,has been deposited at the ECACC on Jun. 13, 1997 under accession No.V97061302.

An example of a variant sequence of SEQ ID NO 9 is SEQ ID NO 11. Thelatter sequence was determined on a serum sample of a patient infectedby the strain MP450, while the former sequence was determined onperipheral blood mononuclear cells (PBMC's) taken from the same patient.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above, comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 39 and/or SEQ ID NO 51 or a variantsequence, said variant sequence showing at least 95% homology with SEQID NO 39 and/or SEQ ID NO 51. An HIV-1 group O strain of this type,termed MP539, has been deposited at the ECACC on Jun. 13, 1997 underaccession No. V97061303.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 1 and/or SEQ ID NO 41 or a variantsequence, said variant sequence showing at least 95% homology with SEQID NO 1 and/or SEQ ID NO 41. A strain of this type is termed MP340throughout this invention.

An example of a variant sequence of SEQ ID NO 1 is SEQ ID NO 3. Thelatter sequence was determined on a serum sample of a patient infectedby the strain MP340, while the former sequence was determined onperipheral blood mononuclear cells (PBMC's) taken from the same patient.The nucleic acids represented by SEQ ID NO 1 and SEQ ID NO 3 show 99%homology, and can be said to belong to strains from the samequasi-species.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 13 and/or SEQ ID NO 47 or a variantsequence, said variant sequence showing at least 95% homology with SEQID NO 13 and/or SEQ ID NO 47. A strain of this type is termed MP448throughout this invention.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 17 and/or SEQ ID NO 49 or a variantsequence, said variant sequence showing at least 95% homology with SEQID NO 17 and/or SEQ ID NO 49. A strain of this type is termed 189throughout this invention.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 19 or a variant sequence showing atleast 95% homology with SEQ ID NO 19. A strain of this type is termed320 throughout this invention.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 21 or a variant sequence showing atleast 95% homology with SEQ ID NO 21. A strain of this type is termedBSD422 throughout this invention.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 23 or a variant sequence showing atleast 95% homology with SEQ ID NO 23. A strain of this type is termedKGT008 throughout this invention.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 25 or a variant sequence showing atleast 95% homology with SEQ ID NO 25. A strain of this type, termedMP575, has been deposited at the ECACC on Jul. 13, 1998, underprovisional accession No. V98071301.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 27 or a variant sequence showing atleast 95% homology with SEQ ID NO 27. A strain of this type is termedBSD189 throughout this invention.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 29 or a variant sequence showing atleast 95% homology with SEQ ID NO 29. A strain of this type is termedBSD649 throughout this invention.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 31 or a variant sequence showing atleast 95% homology with SEQ ID NO 31. A strain of this type is termedBSD242 throughout this invention.

Furthermore, the invention also relates to a strain of HIV-1 group-O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 33 or a variant sequence showing atleast 95% homology with SEQ ID NO 33. A strain of this type is termed533 throughout this invention.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 35 or a variant sequence showing atleast 95% homology with SEQ ID NO 35. A strain of this type is termed772. P94 throughout this invention.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 37 or a variant sequence showing atleast 95% homology with SEQ ID NO 37. A strain of this type is termedMP95B throughout this invention.

Furthermore, the invention also relates to a strain of HIV-1 group O asdefined above comprising in its genome the RNA equivalent of the DNAsequence represented by SEQ ID NO 106 or a variant sequence showing atleast 95% homology with SEQ ID NO 106. A strain of this type, termedMP645, has been deposited at the ECACC on Jul. 13, 1998, underprovisional accession No. V98071302.

Another embodiment of the current invention provides for a nucleic acidmolecule isolated from any of the HIV-1 group O strains as definedabove.

In addition, the current invention provides for an antigen or antigenfragment isolated from any of the HIV-1 group O strains as definedabove.

It is to be understood that the current invention also provides fornucleic acid sequences and antigen sequences which are contained in theabove-mentioned new HIV-1 group O viral strains, and which extend beyondthe explicitly cited sequences represented by SEQ ID NO 1 to 102, 106,135 to 138. The person skilled in the art will realize that, startingfrom the partial sequences disclosed above, it is perfectly possible toobtain the complete genomic information of the respective viruses, bystandard cloning methods such as the construction of a cDNA library orthe construction of a genomic library or by the technique of thepolymerase chain reaction. Sometimes a combination of these methods maybe necessary to obtain the sequence of the full genome.

The following describes the strategies which may be followed to obtainadditional genomic sequence information on HIV-1 group O strains, ofwhich partial sequences have been disclosed above.

1. Construction of a cDNA Library

HIV-1 group O viruses are propagated and isolated using standard methodse.g. by cultivation of peripheral blood lymphocytes (PBMC) from theHIV-infected individual together with stimulated lymphocytes fromhealthy donors, or alternatively by infecting cell lines with the virusin a permanent way. Once virus is detected in the culture supernatantusing standard techniques (e.g. measuring reverse transcriptaseactivity; measuring p24 antigen . . . ), virus is harvested from theculture supernantant by centrifugation under conditions where the virusis pelleted. RNA is obtained by disrupting the virus in a buffercontaining 6M guanidinium chloride and the RNA is pelleted through a5.5M CsCl cushion. The RNA which is resuspended in a suitable buffer isthen phenolized and precipitated with e.g. ethanol and lithium chloride.

cDNA synthesis is performed on the complete RNA or part of the RNA usingcommercially available kits. OligodT primers, random primers, or HIV-1specific primers may be used to prime the cDNA synthesis which is doneby a reverse transcriptase (RT) enzyme. This leads to a first DNA strandwhich is complementary to the initial RNA strand and which formsRNA::DNA hybrids. The RNA strand is removed with Rnase H and the secondDNA strand is then synthesised with DNA polymerase I. The overhangingsingle stranded cDNA ends are removed with T4 DNA polymerase. Theresulting cDNA is ligated to linkers which contain an appropriaterestriction site. After hydrolysis of the cDNA with the appropriaterestriction enzyme, the cDNA of suitable size is isolated (e.g. fromagarose gel after electrophoresis) and ligated in a suitable vector. Thevector containing the cDNA fragments can be propagated in competent E.coli cells using standard methods.

Various techniques to screen for colonies containing HIV-1 specificsequences are known in the art. They involve screening of e.g. a cDNAexpression library (e.g. λgt11) with serum (polyclonal or monoclonalserum) or the screening of a cDNA library with ³²P labelled HIV-1 DNAfragments under non-stringent or stringent hybridization conditions.Background signals are lowered by washing the filters subsequently undermore stringent conditions. After identification of the E. colicontaining the suitable fragment, the fragment is isolated from theplasmid and is introduced (as a complete entity or a fragment thereof)in expression vectors. Using standard techniques, these vectors producethe protein(s) encoded by the inserted DNA fragment. The resultingproteins is further purified and used for the development of diagnosticassays. Sequence information of the virus is obtained from the plasmidcontaining viral DNA sequences.

2. Construction of a Genomic Library

Chromosomal DNA is prepared from cells infected with the HIV-1 group Ovirus (e.g. cells permanently producing the virus) using standardtechniques (Maniatis et al. 1982). This DNA may be used to construct agenomic library (Zabarousky and Allikmets 1986). The chromosomal DNAwhich contains the proviral HIV-1 group O DNA is partially digested witha selected restriction enzyme. Fragments between 9 Kb and 23 Kb,isolated on a 40%-10% sucrose gradient, are manipulated according tostandard techniques in order to introduce them in a vector systemsuitable for the cloning of long DNA fragments e.g. lambda derivedvectors or cosmids.

The vector with the DNA fragment is introduced in a suitable E. colistrain and is further propagated onto plates. Plaques or colonies fromthe genomic library are transferred to nylon or nitrocellulose membranesand screened with enzyme or ³²P labelled DNA fragments of the viralgenome (plaque or colony screening) under non-stringent or stringenthybridization conditions. Colonies or plaques displaying positivesignals are purified from other colonies or plaques. The viral DNA isfurther subcloned and sequenced. Genes or fragments of genes are furthermanipulated using standard techniques in order to express importantviral proteins or epitopes which may be used for the development ofdiagnostic assays.

3. Polymerase Chain Reaction (PCR)

HIV-1 group O viral DNA fragments may also be obtained using thepolymerase chain reaction (PCR) (Kwok et al. 1987) which is a standardtechnology used for the cloning of DNA fragments. PCR may be performedon cellular DNA of cells infected with the virus or on cDNA obtainedfrom viral RNA derived from virus culture, lymphocytes, serum, plasma, .. . . The PCR may use primers which contain specific sequences of thevirus based on sequences of the virus which are already known, oralternatively, primers which contain sequences derived from relatedviruses in regions known to be conserved or not conserved among HIVvariants. Annealing conditions of the primers should preferentially notbe too stringent (e.g. Tm−20° C.), however the best conditions should beexperimentally established. The resulting amplification product issubsequently sequenced and new primers are designed based on the newlygenerated sequence in order to further amplify the viral DNA, againeventually in combination with primers derived from the partiallydetermined sequence of the isolate or from the sequence of relatedviruses.

Example 4 provides the cloning and sequencing strategy followed in orderto obtain the polynucleic acid sequences encoding the antigens Vif, Pol,Vpr, Tat (1st exon), Vpu, Rev (1st exon) and gp160, or fragmentsthereof, from the HIV-1 group O viruses FABA (MP331), MP448, MP539 andMP 645.

Therefore, in addition to the gp41 and V3 sequences described above, thepresent invention further provides for a polynucleic acid containing apolynucleic acid sequence encoding at least part of the Vif, Pol, Vpr,Tat (1st exon), Vpu, Rev (1st exon) or gp160 antigens from any of thefollowing HIV-1 group O strains of the invention: MP340, FABA(MP331),MP450, MP448, 189, MP539, 320, BSD422, KGT008, MP575, BSD189, BSD649,BSD242, 533, 772P94, MP95B and MP645.

The terms “Vif, Pol, Vpr, Tat (1st exon), Vpu, Rev (1st exon) and gp160”are terms for HIV-antigens familiar to the person skilled in the art.Vif and Vpu have important roles during virion morphogenesis. Vif isrequired for the production of fully infectious viruses, while Vpu isnecessary for the efficient release of virus particles budding from thecell membrane in cultured cells (Göttlinger et al, 1993). Vpu alsomediates the rapid degradation of the CD4 receptor molecule in theendoplasmatic reticulum (Willey et al, 1992, Bour et al, 1995). The Vprprotein is involved in the nuclear migration of the prointegrationcomplex (Heinzinger et al, 1994) and is also found in mature virions andhence a structural component of the virus (Paxton et al, 1993). Tat(transactivator) and Rev (regulator of virion expression) are encoded inoverlapping reading frames which generate small regulatory proteinstranslated from multiple spliced mRNAs (Salfeld et al, 1990; Solomin etal, 1990, Furtado et al, 1991). Both proteins are essential for virusreplication and are positive regulators of gene expression (Arya et al,1985; Feinberg et al, 1986). Pol is encoded by the coding gene pol,which overlaps with the gag information but in a different readingframe. Pol is a precursor protein which is autocleaved to form thefollowing viral enzymes: a protease, a reverse transcriptase withpolymerase activity and Rnase H activity, and an integrase (Ross et al,1991). FIG. 8 illustrates the sequence of a large genomic fragment froma number of HIV-1 group O strains of the current invention (MP645 (SEQID NO 106), MP331 (SEQ ID NO 103), MP448 (SEQ ID NO 104) and MP539(SEQID NO 105)), and the location of the Vif, Pol, Vpr, Tat (1st exon), Vpu,Rev (1st exon) and gp160 (partially) genes in these sequences.

The present invention thus provides for a polynucleic acid comprising anucleotide sequence chosen from the group of

(I) a nucleotide sequence represented by any of SEQ ID NO 103, SEQ ID NO104, SEQ ID NO 105, SEQ ID NO 106, or

(ii) a nucleotide sequence complementary to a sequence according to (I),or

(iii) a nucleotide sequence showing at least 95%, preferably 96%, 97%,98% and most preferably 99% homology to a sequence according to (I), or

(iv) a nucleotide sequence according to (I) whereby T is replaced by U,or

(v) a nucleotide sequence according to (I) whereby at least onenucleotide is substituted by a nucleotide analogue, or

(vi) a fragment a sequence of at least 15, preferably 16, 17, 18, 19,20, 21, 22, 23, 24, 25 up to 50 contiguous nucleotides of any of thenucleotide sequences according to (I) to (v), and characterized by thefact that it selectively hybridizes to the polynucleic acid from whichit is derived, and/or selectively amplifies the polynucleic acid fromwhich it is derived.

More particularly, the present invention provides for a polynucleic acidconsisting of a nucleotide sequence chosen from the group of

(I) a nucleotide sequence represented by any of SEQ ID NO 103, SEQ ID NO104, SEQ ID NO 105, SEQ ID NO 106, or

(ii) a nucleotide sequence complementary to a sequence according to (I),or

(iii) a nucleotide sequence showing at least 95%, preferably 96%, 97%,98% and most preferably 99% homology to a sequence according to (I), or

(iv) a nucleotide sequence according to (I) whereby T is replaced by U,or

(v) a nucleotide sequence according to (I) whereby at least onenucleotide is substituted by a nucleotide analogue

(vi) a fragment a sequence of at least 15, preferably 16, 17, 18, 19,20, 21, 22, 23, 24, 25 up to 50 contiguous nucleotides of any of thenucleotide sequences according to (I) to (v), and characterized by thefact that it selectively hybridizes to the polynucleic acid from whichit is derived, and/or selectively amplifies the polynucleic acid fromwhich it is derived.

The above-described nucleic acid fragment may be used as a specifichybridization probe for the detection of the nucleic acids of thecurrent invention.

The term “selectively hybridizing” means that the hybridization signalobtained after hybridization of the fragment with the nucleic acid fromwhich it is derived, is more intense than the hybridization signalobtained when the fragment is hybridized to the corresponding nucleicacid from another HIV-1 group O strain, under the same stringenthybridization and wash conditions. In practice this means that thenucleic acid fragment will show at least one mismatched nucleotide withthe sequence of the corresponding nucleic acid fragment of another HIV-1group O strain.

The term “stringent hybridization conditions” implies that thehybridization takes place at a temperature which is situatedapproximately between Tm and (Tm−10° C.), whereby Tm represents thecalculated melting temperature of the target nucleic. It is generallyknown that the stringency depends on the percentage mismatches(=non-matching nucleotides upon alignment) present in the hybridizingduplex. According to a simplified formula, the hybridization temperaturemay be calculated as follows: Tm−1.2 (% mismatch). A temperaturedecrease of 10° C. implies a maximum percentage of allowed mismatches of8.3%.

The nucleic acid fragment as described above may also be used as aspecific amplification primer of the nucleic acids of the currentinvention.

The term “selective amplification” refers to the fact that said nucleicacid fragment may initiate a specific amplification reaction of thenucleic acids of the invention (e.g. a polymerase chain reaction) in thepresence of other nucleic acids, under appropriate amplificationconditions. It means that, under the appropriate amplificationconditions, only the nucleic acids of the invention will be amplified,and not the other nucleic acids possibly present.

Preferred embodiments of the invention comprise polynucleic acids orfragments thereof. as specified below.

A polynucleic acid comprising SEQ ID NO 103, or comprising a fragmentconsisting of at least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23,24, 25, up to 50 contiguous nucleotides of said polynucleic acid, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 103.

A polynucleic acid comprising SEQ ID NO 104, or comprising a fragmentconsisting of at least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23,24, 25, up to 50 contiguous nucleotides of said polynucleic acid, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 104.

A polynucleic acid comprising SEQ ID NO 105, or comprising a fragmentconsisting of at least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23,24, 25, up to 50 contiguous nucleotides of said polynucleic acid, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 105.

A polynucleic acid comprising SEQ ID NO 106, or comprising a fragmentconsisting of at least 15, preferably 16, 17, 18, 19, 20, 21, 22, 23,24, 25, up to 50 contiguous nucleotides of said polynucleic acid, saidfragment characterized by the fact that it selectively hybridizes to thepolynucleic acid comprising SEQ ID NO 106.

A polynucleic acid fragment consisting of a sequence of at least 15,preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, up to 50 contiguousnucleotides of SEQ ID NO 103, said fragment characterized by the factthat it selectively amplifies the polynucleic acid from which it isderived.

A polynucleic acid fragment consisting of a sequence of at least 15,preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, up to 50 contiguousnucleotides of SEQ ID NO 104, said fragment characterized by the factthat it selectively amplifies the polynucleic acid from which it isderived.

A polynucleic acid fragment consisting of a sequence of at least 15,preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, up to 50 contiguousnucleotides of SEQ ID NO 105, said fragment characterized by the factthat it selectively amplifies the polynucleic acid from which it isderived.

A polynucleic acid fragment consisting of a sequence of at least 15,preferably 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, up to 50 contiguousnucleotides of SEQ ID NO 106, said fragment characterized by the factthat it selectively amplifies the polynucleic acid from which it isderived.

The invention further provides for an antigen comprising at least partof Vif, Pol, Vpr, Tat (1st exon), Vpu, Rev (1st exon) and/or gp160encoded by the nucleic acid sequences as described above from any of thefollowing HIV-1 group O strains: MP340, FABA(MP331), MP450, MP448, 189,MP539, 320, BSD422, KGT008, MP575, BSD189, BSD649, BSD242, 533, 772P94,MP95B and MP645.

The current invention more particularly provides for an antigencomprising at least one amino acid sequence chosen from the followinggroups of sequences

Ii) an amino acid sequence represented by any of SEQ ID NO 107, SEQ IDNO 108, SEQ ID NO 109 and SEQ ID NO 110 representing the Vif antigen,

(ii) an amino acid sequence represented by any of SEQ ID NO 111, SEQ IDNO 112, SEQ ID NO 113 and SEQ ID NO 114 representing the Vpu antigen,

(iii) an amino acid sequence represented by any of SEQ ID NO 115, SEQ IDNO 116, SEQ ID NO 117 and SEQ ID NO 118 representing the Vpr antigen,

(iv) an amino acid sequence represented by any of SEQ ID NO 119, SEQ IDNO 120, SEQ ID NO 121 and SEQ ID NO 122 representing the Tat antigen,

(v) an amino acid sequence represented by any of SEQ ID NO 123, SEQ IDNO 124, SEQ ID NO 125 and SEQ ID NO 126 representing the Rev antigen,

(vi) an amino acid sequence represented by any of SEQ ID NO 127, SEQ IDNO 128, SEQ ID NO 129 and SEQ ID NO 130 representing the Pol antigen,

(vii) an amino acid sequence represented by any of SEQ ID NO 2, SEQ IDNO 4, SEQ ID NO 6, SEQ ID NO 8, SEQ ID NO 10, SEQ ID NO 12, SEQ ID NO14, SEQ ID NO 16, SEQ ID NO 18, SEQ ID NO 2, SEQ ID NO 22, SEQ ID NO 24,SEQ ID NO 26, SEQ ID NO 28, SEQ ID NO 30, SEQ ID NO 32, SEQ ID NO 34,SEQ ID NO 36, SEQ ID NO 38, SEQ ID NO 40, SEQ ID NO 42, SEQ ID NO 44,SEQ ID NO 46, SEQ ID NO 48, SEQ ID NO 50, SEQ ID NO 52, SEQ ID NO 132,and SEQ ID NO 134 representing at least part of the Env antigen, or

(viii) a fragment of any of the above-mentioned antigens Ii) to (vii),said fragment consisting of at least 8, preferably 9, 10, 11,12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50 up to the maximum numberof contiguous amino acids of the amino acid sequence of said antigen,and being characterized by the fact that it specifically reacts withantibodies raised against said antigen.

The current invention further provides for an antigen consisting of anamino acid sequence chosen from the following groups of sequences:

Ii) an amino acid sequence SEQ ID NO 107, SEQ ID NO 108, SEQ ID NO 109and SEQ ID NO 110 representing the Vif antigen,

(i) an amino acid sequence SEQ ID NO 111, SEQ ID NO 112, SEQ ID NO 113and SEQ ID NO 114 representing the Vpu antigen,

(iii) an amino acid sequence SEQ ID NO 115, SEQ ID NO 116, SEQ ID NO 117and SEQ ID NO 118 representing the Vpr antigen,

(iv) an amino acid sequence SEQ ID NO 119, SEQ ID NO 120, SEQ ID NO 121and SEQ ID NO 122 representing the Tat antigen,

(v) an amino acid sequence SEQ ID NO 123, SEQ ID NO 124, SEQ ID NO 125and SEQ ID NO 126 representing the Rev antigen,

(vi) an amino acid sequence SEQ ID NO 127, SEQ ID NO 128, SEQ ID NO 129and SEQ ID NO 130 representing the Pol antigen,

(vii) an amino acid sequence represented by any of SEQ ID NO 2, SEQ IDNO 4, SEQ ID NO 6, SEQ ID NO 8, SEQ ID NO 10, SEQ ID NO 12, SEQ ID NO14, SEQ ID NO 16, SEQ ID NO 18, SEQ ID NO 20, SEQ ID NO 22, SEQ ID NO24, SEQ ID NO 26, SEQ ID NO 28, SEQ ID NO 30, SEQ ID NO 32, SEQ ID NO34, SEQ ID NO 36, SEQ ID NO 38, SEQ ID NO 40, SEQ ID NO 42, SEQ ID NO44, SEQ ID NO 46, SEQ ID NO 48, SEQ ID NO 50, SEQ ID NO 52, SEQ ID NO132, and SEQ ID NO 134 representing at least part of the Env antigen, or

(viii) a fragment of any of the above-mentioned antigens (I) to (vii),said fragment consisting of at least 8, preferably 9, 10,11,12, 13, 14,15, 16, 17, 18, 19, 20, 21,22, 23, 24, 25, 50 up to the maximum numberof contiguous amino acids of the amino acid sequence of said antigen,and being characterized by the fact that it specifically reacts withantibodies raised against said antigen.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain MP340 comprising at least one of the sequencesaccording to SEQ ID NO 2, 4, 42, 59, 60, 61, 73, 85, 86, 100, orfragments thereof with said fragments specifically reacting withantibodies raised against the antigen they are derived from;

(ii) a polynucleic acid encoding an antigen according to (I) andcomprising at least one of the nucleotide sequences according to SEQ IDNO 1, 3, 41, including homologous sequences, complementary sequences,and fragments hybridizing thereto;

(iii) a virus strain comprising in its genome a polynucleic acidaccording to (ii), more particularly a virus strain termed MP340, aswell as polynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain MP331 (or FABA) comprising at least one of thesequences according to SEQ ID NO 6, 8, 44, 56, 57, 58, 82, 83, 84, 138or fragments thereof with said fragments specifically reacting withantibodies raised against the antigen they are derived from;

(ii) an antigen derived from the Vif, Vpu, Vpr, Tat, Rev, Pol and Envprotein of the new HIV-1 group O strain MP331 (or FABA) comprising atleast one of the sequences according to SEQ ID NO 107, 111, 115, 119,123, 127, 131, or fragments thereof with said fragments specificallyreacting with antibodies raised against the antigen they are derivedfrom;

(iii) a polynucleic acid encoding an antigen according to (I) or (ii)and comprising at least one of the nucleic acid sequences according toSEQ ID NO 5, 7, 43, 103, including homologous sequences, complementarysequences, and fragments hybridizing thereto;

(iv) a virus strain comprising in its genome said a polynucleic acidaccording to (iii), more particularly a virus strain termed MP331 (FABA)deposited at the ECACC under accession number V97061301, as well aspolynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain MP450 comprising at least one of the sequencesaccording to SEQ ID NO 10, 12, 46, 62, 63, 64, 73, 87, 100, or fragmentsthereof with said fragments specifically reacting with antibodies raisedagainst the antigen they are derived from;

(ii) a polynucleic acid encoding an antigen according to (I) andcomprising at least one of the nucleotide sequences according to SEQ IDNO 9, 11, 45, including homologous sequences, complementary sequences,and fragments hybridizing thereto;

(iii) a virus strain comprising in its genome a polynucleic acidaccording to (ii), more particularly a virus strain termed MP450deposited at the ECACC under accession number V97061302, as well aspolynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain MP448 comprising at least one of the sequencesaccording to SEQ ID NO 14, 16, 48, 65, 66, 67, 76, 88, 101, or fragmentsthereof with said fragments specifically reacting with antibodies raisedagainst the antigen they are derived from;

(ii) an antigen derived from the Vif, Vpu, Vpr, Tat, Rev, Pol and Envprotein of the new HIV-1 group O strain MP448 comprising at least one ofthe sequences according to SEQ ID NO 108, 112, 116, 120, 124, 128, 132or fragments thereof with said fragments specifically reacting withantibodies raised against the antigen they are derived from;

(iii) a polynucleic acid encoding an antigen according to (I) or (ii)and comprising at least one of the nucleic acid sequences according toSEQ ID NO 13, 15, 47, 104, including homologous sequences, complementarysequences, and fragments hybridizing thereto;

(iv) A virus strain comprising in its genome said polynucleic acidaccording to (iii), more particularly a virus strain termed MP448, aswell as polynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain 189 comprising at least one of the sequencesaccording to SEQ ID NO 18, 50, 53, 54, 55, 73, 90, 91, or fragmentsthereof with said fragments specifically reacting with antibodies raisedagainst the antigen they are derived from;

(ii) a polynucleic acid encoding an antigen according to (I) andcomprising at least one of the nucleotide sequences according to SEQ IDNO 17, 49, including homologous sequences, complementary sequences, andfragments hybridizing thereto,

(iii) a virus strain comprising in its genome a polynucleic acidaccording to (ii), more particularly a virus strain termed 189, as wellas polynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain MP539 comprising at least one of the sequencesaccording to SEQ ID NO 40, 52, 68, 69, 70, 71, 89, 95 or fragmentsthereof with said fragments specifically reacting with antibodies raisedagainst the antigen they are derived from;

(ii) an antigen derived from the Vif, Vpu, Vpr, Tat, Rev, Pol and Envprotein of the new HIV-1 group O strain MP539 comprising at least one ofthe sequences according to SEQ ID NO 109, 113, 117, 121, 125, 129, 133,or fragments thereof with said fragments specifically reacting withantibodies raised against the antigen they are derived from.

(iii) a polynucleic acid encoding an antigen according to (I) or (ii)and comprising at least one of the nucleic acid sequences according toSEQ ID NO 39, 51, 105, including homologous sequences, complementarysequences, and fragments hybridizing thereto,

(iv) A virus strain comprising in its genome said polynucleic acidaccording to (iii), more particularly a virus strain deposited at theECACC under accession number V97061303, as well as polynucleic acids andantigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain 320 comprising at least one of the sequencesaccording to SEQ ID NO 20, 92, or fragments thereof with said fragmentsspecifically reacting with antibodies raised against the antigen theyare derived from;

(ii) a polynucleic acid encoding an antigen according to (I) andcomprising the nucleotide sequence according to SEQ ID NO 19 includinghomologous sequences, complementary sequences, and fragments hybridizingthereto;

(iii) a virus strain comprising in its genome a polynucleic acidaccording to (ii), more particularly a virus strain termed 320, as wellas polynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain BSD422 comprising at least one of the sequencesaccording to SEQ ID NO 22, 80, 79, or fragments thereof with saidfragments specifically reacting with antibodies raised against theantigen they are derived from;

(ii) a polynucleic acid encoding an antigen according to (I) andcomprising the nucleotide sequence according to SEQ ID NO 21 includinghomologous sequences, complementary sequences, and fragments hybridizingthereto;

(iii) a virus strain comprising in its genome a polynucleic acidaccording to (ii), more particularly a virus strain termed BSD422, aswell as polynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain KGT008 comprising at least one of the sequencesaccording to SEQ ID NO 24, 79, 99? or fragments thereof with saidfragments specifically reacting with antibodies raised against theantigen they are derived from;

(ii) a polynucleic acid encoding an antigen according to (I) andcomprising the nucleotide sequence according to SEQ ID NO 23 includinghomologous sequences, complementary sequences, and fragments hybridizingthereto;

(iii) a virus strain comprising in its genome a polynucleic acidaccording to (ii), more particularly a virus strain termed KGT008, aswell as polynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain MP575 comprising the sequence according to SEQ IDNO 26, or fragments thereof with said fragments specifically reactingwith antibodies raised against the antigen they are derived from;

(ii) a polynucleic acid encoding an antigen according to Ii) andcomprising the nucleotide sequence according to SEQ ID NO 25 includinghomologous sequences, complementary sequences, and fragments hybridizingthereto;

(iii) A virus strain comprising in its genome said polynucleic acidaccording to (ii), more particularly a virus strain deposited at theECACC under provisional accession number V98071301, as well aspolynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain BSD189 comprising at least one of the sequencesaccording to SEQ ID NO 28, 72, 95, or fragments thereof with saidfragments specifically reacting with antibodies raised against theantigen they are derived from;

(ii) a polynucleic acid encoding an antigen according to (I) andcomprising the nucleotide sequence according to SEQ ID NO 27 includinghomologous sequences, complementary sequences, and fragments hybridizingthereto;

(iii) a virus strain comprising in its genome a polynucleic acidaccording to (ii), more particularly a virus strain termed BSD189, aswell as polynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain BSD649 comprising at least one of the sequencesaccording to SEQ ID NO 30, 77, 98, or fragments thereof with saidfragments specifically reacting with antibodies raised against theantigen they are derived from;

(ii) a polynucleic acid encoding an antigen according to (I) andcomprising the nucleotide sequence according to SEQ ID NO 29 includinghomologous sequences, complementary sequences, and fragments hybridizingthereto;

(iii) a virus strain comprising in its genome a polynucleic acidaccording to (ii), more particularly a virus strain termed BSD649, aswell as polynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain BSD242 comprising at least one of the sequencesaccording to SEQ ID NO 32, 81, 96, or fragments thereof with saidfragments specifically reacting with antibodies raised against theantigen they are derived from;

(ii) a polynucleic acid encoding an antigen according to (I) andcomprising the nucleotide sequence according to SEQ ID NO 31 includinghomologous sequences, complementary sequences, and fragments hybridizingthereto;

(iii) a virus strain comprising in its genome a polynucleic acidaccording to (ii), more particularly a virus strain termed BSD242, aswell as polynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain 533 comprising at least one of the sequencesaccording to SEQ ID NO 34, 81, 93, or fragments thereof with saidfragments specifically reacting with antibodies raised against theantigen they are derived from;

(ii) a polynucleic acid encoding an antigen according to (I) andcomprising the nucleotide sequence according to SEQ ID NO 33 includinghomologous sequences, complementary sequences, and fragments hybridizingthereto;

(iii) a virus strain comprising in its genome a polynucleic acidaccording to (ii), more particularly a virus strain termed 533, as wellas polynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain 772P94 comprising at least one of the sequencesaccording to SEQ ID NO 36, 75, 94, or fragments thereof with saidfragments specifically reacting with antibodies raised against theantigen they are derived from;

(ii) a polynucleic acid encoding an antigen according to (I) andcomprising the nucleotide sequence according to SEQ ID NO 35 includinghomologous sequences, complementary sequences, and fragments hybridizingthereto;

(iii) a virus strain comprising in its genome a polynucleic acidaccording to (ii), more particularly a virus strain termed 772P94, aswell as polynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O strain MP95B comprising at least one of the sequencesaccording to SEQ ID NO 38, 74, 102, or fragments thereof with saidfragments specifically reacting with antibodies raised against theantigen they are derived from;

(ii) a polynucleic acid encoding an antigen according to (I) andcomprising the nucleotide sequence according to SEQ ID NO 37 includinghomologous sequences, complementary sequences, and fragments hybridizingthereto;

(iii) a virus strain comprising in its genome a polynucleic acidaccording to (ii), more particularly a virus strain termed MP95B, aswell as polynucleic acids and antigens isolated therefrom.

The current invention thus also relates to:

(I) an antigen derived from the gp160 env precursor protein of the newHIV-1 group O stain MP645 comprising at least one of the sequencesaccording to SEQ ID NO 135, 136, 137, or fragments thereof with saidfragments specifically reacting with antibodies raised against theantigen they are derived from;

(ii) an antigen derived from the Vif, Vpu, Vpr, Tat, Rev, Pol and Envprotein of the new HIV-1 group O strain MP645 comprising one of thesequences according to SEQ ID NO 110, 114, 118, 122, 126, 130, 134, orfragments thereof with said fragments specifically reacting withantibodies raised against the antigen they are derived from.

(iii) a polynucleic acid encoding an antigen according to (I) or (ii)and comprising the nucleic acid sequence according to SEQ ID NO 106,including homologous sequences, complementary sequences, and fragmentshybridizing thereto;

(iv) A virus strain comprising in its genome said polynucleic acidaccording to (iii), more particularly a virus strain deposited at theECACC under provisional accession number V98071302, as well aspolynucleic acids and antigens isolated therefrom.

In another embodiment, the invention provides for an antibody,preferably a monoclonal antibody, raised against an antigen or antigenfragment as described above. Such an antibody recognizes specificallythe antigen or the antigen fragment to which it has been raised.

According to an alternative embodiment, the present invention alsorelates to an antigen-binding fragment of the antibody, said fragmentbeing of the F(ab′)₂, Fab or single chain Fv type, or any type ofrecombinant antibody derived from said specific antibodies or monoclonalantibodies, provided that said antibody fragment or recombinant antibodystill recognizes specifically the antigen or antigen fragment to whichit has been raised.

The expression “antibody recognizing specifically” means that thebinding between the antigen as a ligand and a molecule containing anantibody combining site, such as a Fab portion of a whole antibody, isspecific, signifying that no cross-reaction occurs.

The expression “antibody specifically raised against a compound” meansthat the sole immunogen used to produce said antibody was said compound.

The possible cross-reactivity of polyclonal antisera may be eliminatedby preabsorption of the polyclonal antiserum against the cross-reactingantigenic determinants.

In a preferential embodiment, the above-mentioned antibodies areneutralizing antibodies, i.e. antibodies capable of in vitro inhibitionof viral growth, determined according to methods known in the art.

Neutralizing antibodies may be used as a reagent in a so-called “passivevaccine” composition, i.e. a composition conferring temporary protectionagainst an infection, upon injection in an individual. The inventionalso relates to passive vaccine compositions, comprising any of theabove-mentioned neutralizing antibodies.

The monoclonal antibodies of the invention can be produced by anyhybridoma liable to be formed according to classical methods fromsplenic cells of an animal, particularly of a mouse or rat, immunizedwith the antigen of the invention, defined above on the one hand, and ofcells of a myeloma cell line on the other hand, and to be selected bythe ability of the hybridoma to produce the monoclonal antibodiesrecognizing the antigen which has been initially used for theimmunization of the animals.

The monoclonal antibodies according to a preferred embodiment of theinvention may be humanized versions of the mouse monoclonal antibodiesmade by means of recombinant DNA technology, departing from the mouseand/or human genomic DNA sequences coding for H and L chains or fromcDNA clones coding for H and L chains.

Also fragments derived from these monoclonal antibodies such as Fab,F(ab)′₂ and ssFv (“single chain variable fragment”), providing they haveretained the original binding properties, form part of the presentinvention. Such fragments are commonly generated by, for instance,enzymatic digestion of the antibodies with papain, pepsin, or otherproteases. It is well known to the person skilled in the art thatmonoclonal antibodies, or fragments thereof, can be modified for varioususes.

The antibodies involved in the invention can be labelled by anappropriate label of the enzymatic, fluorescent, or radioactive type.

The invention also relates to the use of the antigens of the invention,or fragments thereof, for the selection of recombinant antibodies by theprocess of repertoire cloning (Perrson et al., 1991).

The present invention further relates to an anti-idiotype antibodyraised against any of the antibodies as defined above.

The term “anti-idiotype antibodies” refers to monoclonal antibodiesraised against the antigenic determinants of the variable region ofmonoclonal antibodies themselves raised against the antigens of theinvention. These antigenic determinants of immunoglobulins are known asidiotypes (sets of idiotopes) and can therefore be considered to be the“fingerprint” of an antibody (for review see de Préval, 1978; Fleishmannand Davie,1984). The methods for production of monoclonal anti-idiotypicantibodies have been described by Gheuens and McFarlin (1982).Monoclonal anti-idiotypic antibodies have the property of forming animmunological complex with the idiotype of the monoclonal antibodyagainst which they were raised. In this respect the monoclonal antibodyis often referred to as Ab1, and the anti-idiotypic antibody is referredto as Ab2. These anti-idiotype Ab2s may be used as substitutes for thepolypeptides of the invention or as competitors for binding of thepolypeptides of the invention to their target.

The present invention further relates to antisense peptides derived fromthe antigens of the invention as described above.

More particularly, the term “antisense peptide” is reviewed by Blalock(1990) and by Roubos (1990). In this respect, the molecular recognitiontheory (Blalock, 1990) states that not only the complementary nucleicacid sequences interact but that, in addition, interacting sites inproteins are composed of complementary amino acid sequences (senseligand with receptor or sense ligand with antisense peptides). Thus, twopeptides derived from complementary nucleic acid sequences in the samereading frame will show a total interchange of their hydrophobic andhydrophilic amino acids when the amino terminus of one is aligned withthe carboxy terminus of the other. This inverted hydropathic patternmight allow two such peptides to assume complementary conformationsresponsible for specific interaction.

The antisense peptides can be prepared as described in Ghiso et al.(1990). By means of this technology it is possible to logicallyconstruct a peptide having a physiologically relevant interaction with aknown peptide by simple nucleotide sequence analysis forcomplementarity, and synthesize the peptide complementary to the bindingsite.

The present invention further relates to a diagnostic method fordetecting the presence of an HIV-1 infection, said method comprising

the detection of antibodies against HIV-1, including HIV-group O, usingany of the antigens or antigen fragments of the invention as describedabove, and/or

the detection of viral antigen originating from HIV-1, including HIV-1group O, using any of the antibodies of the invention as described aboveand/or

the detection of viral nucleic acids originating from HIV-1, includingHIV-1 group O, using any of the nucleic acids or nucleic acid fragmentsof the invention as described above, in a biological sample.

Preferably the above-mentioned diagnostic method for detecting thepresence of an HIV-1 infection also includes the detection of an HIV-1group O infection, and more particularly also includes the detection ofan infection caused by any of the HIV-1 group O strains of the currentinvention.

The term “biological sample” refers to any biological sample (tissue orfluid) possibly containing HIV nucleic acids, and/or HIV antigens and/orantibodies against HIV, and refers more particularly to blood, serum,plasma, organs or tissue samples.

In most instances, the [HIV-1 group O]-reagents (=antigens and/orantibodies and/or nucleic acids) of the invention will be used inmethods which combine them with other HIV-reagents (=antigens and/orantibodies and/or nucleic acids). The addition of the HIV-1 type Oreagents of the current invention to methods and kits for detection ofHIV-infection in general, may result in methods and kits showing

a higher sensitivity, and/or

a higher discriminating power between different types of HIV-infection,for example HIV-1 group M, HIV-1 group O and HIV-2 infection.

The term “sensitivity” refers to the ratio of positively reactingsamples/the number of truly infected samples.

More specifically, the present invention relates to a method for invitro diagnosis of a HIV-1 infection, including a HIV-1 group Oinfection, comprising at least the step of contacting a biologicalsample with:

a HIV-1 group O antigen, or antigen fragment, as defined above, underconditions allowing the formation of an immunological complex, and/or,

a HIV-1 group O nucleic acid, or nucleic acid fragment, as definedabove, under conditions allowing the formation of a hybridizationcomplex, with the nucleic acids of said sample being possibly amplifiedprior to hybridization, and/or,

an antibody specifically directed against an HIV-1 group O antigen asdefined above, under conditions allowing the formation of animmunological complex, and/or,

an anti-idiotype antibody as defined above, under conditions allowingthe formation of an antibody-anti-idiotypic complex, and/or,

an antisense peptide as defined above, under conditions allowing theformation of an antigen-antisense peptide complex,

and subsequently detecting the complexes formed.

In a more specific embodiment, the invention relates to a method fordetecting the presence of antibodies against HIV-1 in a biologicalsample, in particular antibodies against an HIV-1 group O strain,preferably a serum sample, comprising the following steps:

contacting the biological sample taken from a patient with at least oneantigen or antigen fragment as described above, under conditionsenabling the formation of an immunological complex, and

detecting the immunological complex formed between said antigen orantigen fragment and the antibodies possibly present in the sample.

Conditions allowing the formation of an immunological complex are knownto the person skilled in the art.

In a special embodiment, the antigens being used in the above-describedmethod for detection of anti-HIV-1 group O antibodies, can be replacedby anti-idiotype antibodies as described above, acting as theirequivalents.

Conditions allowing the formation of an antibody-anti-idiotypic complexare known in the art.

The invention farther relates to a method for detecting the presence ofan antigen or an antigen fragment of HIV-1, in particular an antigen orantigen fragment of an HIV-1 group O strain, in a biological samplecomprising the following steps:

contacting the biological sample taken from a patient with at least oneantibody as described above under conditions enabling the formation ofan immunological complex, and

detecting the immunological complex formed between said antibody and theantigen or antigen fragment possibly present in the sample.

In a special embodiment, the antibodies being used in theabove-described method for detection of HIV-1 group O antigens, may bereplaced by anti-sense peptides as described above, acting as theirequivalents.

Conditions allowing the formation of an antigen-antisense peptidecomplex are known in the art.

Design of immunoassays is subject to a great deal of variation, and manyformats are known in the art. Protocols may, for example, use solidsupports, or immunoprecipitation. Most assays involve the use oflabelled antibody or polypeptide; the labels may be, for example,enzymatic, fluorescent, chemoluminescent, radioactive, or dye molecules.Assays which amplify the signals from the immune complex are also known,examples of which are assays which utilize biotin and avidin orstreptavidin, and enzyme-labelled and mediated immunoassays, such asELISA assays.

An advantageous embodiment provides for a method for detection ofanti-HIV-1 group O antibodies in a sample, whereby the antigens orantigen fragments of the invention are immobilized on a solid support,for example on a membrane strip. Different antigens or antigen fragmentsof the invention may be immobilized together or next to each other (e.g.in the form of parallel lines). The antigens of the invention may alsobe combined with other antigens, e.g. antigens from other HIV-1 group Ostrains, or from HIV-1 group M or from HIV-2 strains.

The combination of different antigens in one single detection method asdescribed above has certain advantages, such as:

achieving a higher test sensitivity: e.g. by combining several antigenicdeterminants from different HIV-strains, the total number of positivelyreacting sera originating from HIV-infected patients will be greater,and/or

enabling differentiation between individuals infected by differentstrains of HIV, more particularly enabling differentiation between HIV-1group M, HIV-1 group O and HIV-2 infected patients.

The invention thus also relates to a solid support onto which theantigens of the invention, possibly in combination with other antigens,have been immobilized.

Another embodiment of the invention provides for a method for detectingthe presence of HIV-1 nucleic acids, including HIV-1 group O nucleicacids, in a biological sample, comprising:

(I) possibly extracting the polynucleic acids contained in the sample,

(ii) possibly amplifying the HIV-1 polynucleic acids, including theHIV-1 group O polynucleic acids, with a suitable primer pair,

(iii) detecting the amplified nucleic acids, after hybridization with aprobe as described above.

The expression “a suitable primer pair” refers to a pair of primersallowing the amplification of the target region to which the probes ofthe current invention hybridize. Depending on the application, theprimer sequences may be chosen such that they amplify specifically thenucleic acids of the current invention, or, on the other hand, it may bepreferred to obtain a more general amplification, e.g. of all or nearlyall HIV-1 group O sequences, or of all or nearly all HIV-1 sequences,and even HIV-2 sequences.

In case a general amplification of HIV-1 group M and HIV-1 group Osequences is preferred, the following pair of primers may be used toamplify part of the gp41 region:

5′-GGGTTCTTGGGAGCAGCAGGAAGCACTATGGGCG-3′ (SEQ ID NO 139), and

5′-TCTGAAACGACAGAGGTGAGTATCCCTGCCTAA-3′ (SEQ ID NO 140)).

In case a more specific amplification of the gp41 region of HIV-1 groupO strains is preferred, the following pair of primers may be used:

5′-TGGATCCCACAGTGTACTGAAGGGTATAGTGCA-3′ (SEQ ID NO 141), and

5′-CATTTAGTTATGTCAAGCCAATTCCAAA-3′ (SEQ ID NO 142)).

The invention also relates to a method for genotyping HIV-1 or HIV-1type O strains, comprising the following steps:

possibly extracting the nucleic acids from the sample,

amplifying the HIV-1 or HIV-1 type O nucleic acids using a suitableprimer pair,

hybridizing the nucleic acids of the sample with at least one probe asdescribed above,

detecting the hybrids formed,

inferring the presence of one or more HIV-1 or HIV-1 type O genotypesfrom the hybridization pattern obtained.

The term “genotyping” refers to the typing of HIV-strains according tothe sequence of their nucleic acids. Depending on the application, itmay be the intention of a genotyping assay to differentiate betweenlarge groups of HIV-strains (e.g. HIV-1 group M; HIV-1 group O or HIV-2)or to subtype smaller entities (such as e.g. the clades withing HIV-1group M (A to J)). Subtyping within HIV-1 group O may also beaccomplished using the nucleic acids of the current invention.

Conditions allowing hybridization are known in the art and e.g.exemplified in Maniatis et al. (1982). However, according to thehybridization solution (SSC, SSPE, etc.), the probes used should behybridized at their appropriate temperature in order to attainsufficient specificity (in some cases differences at the level of onenucleotide mutation are to be discriminated).

Amplification of nucleic acids present in a sample prior to detection invitro may be accomplished by first extracting the nucleic acids presentin the sample according to any of the techniques known in the art, andsecond, amplifying the target nucleic acid by any amplification methodas specified above. In case of extraction of RNA, generation of cDNA isnecessary; otherwise cDNA or genomic DNA is extracted.

The term “labelled” refers to the use of labelled nucleic acids. Thismay include the use of labelled nucleotides incorporated during thepolymerase step of the amplification such as illustrated by Saiki et al.(1988) or Bej et al. (1990) or labelled primers, or by any other methodknown to the person skilled in the art. Labels may be isotopic (³²P,³⁵S, etc.) or non-isotopic (biotin, digoxigenin, etc.).

Suitable assay methods for purposes of the present invention to detecthybrids formed between oligonucleotide probes according to the inventionand the nucleic acid sequences in a sample may comprise any of the assayformats known in the art. For example, the detection can be accomplishedusing a dot blot format, the unlabelled amplified sample being bound toa membrane, the membrane being incubated with at least one labelledprobe under suitable hybridization and wash conditions, and the presenceof bound probe being monitored. Probes can be labelled withradioisotopes or with labels allowing chromogenic or chemiluminescentdetection such as horse-radish peroxidase coupled probes.

An alternative is a “reverse” dot-blot format, in which the amplifiedsequence contains a label. In this format, the unlabelledoligonucleotide probes are bound to a solid support and exposed to thelabelled sample under appropriate stringent hybridization and subsequentwashing conditions. It is to be understood that also any other assaymethod which relies on the formation of a hybrid between the nucleicacids of the sample and the oligonucleotide probes according to thepresent invention may be used.

According to an advantageous embodiment, the process of detecting HIV-1type O nucleic acids contained in a biological sample comprises thesteps of contacting amplified copies of the nucleic acids present in thesample, with a solid support on which probes as defined above, have beenpreviously immobilized. Preferably, the amplified nucleic acids arelabelled in order to subsequently detect hybridization.

In a very specific embodiment, the probes have been immobilized on amembrane strip in the form of parallel lines. This type of reversehybridization method is specified further as a Line Probe Assay (LiPA),and has been described more extensively in for example WO 94/12670.

The invention thus also relates to a solid support onto which thenucleic acids of the invention have been immobilized.

The invention also provides for a composition comprising at least one ofthe antigens or antigen fragments as above described, and/or at leastone of the nucleic acids or nucleic acid fragments as above described,and/or an antibody as above described.

Examples of such compositions may be e.g. a diagnostic kit, animmunogenic composition, e.a.

In particular, the invention provides for a kit for the detection of thepresence of an HIV-1 infection, comprising at least one of the antigensor antigen fragments as described above and/or at least one of thenucleic acids or nucleic acid fragments as described above and/or anantibody as described above.

More specifically, the current invention provides for a diagnostic kitfor determining the presence of HIV-1 nucleic acids, including HIV-1type O nucleic acids, in a biological sample, said kit comprising atleast one nucleic acid fragment as described above. This nucleic acidfragment may be used as a primer or a probe in said kit.

In addition, the current invention provides for a kit for genotypingHIV-1 strains, including HIV-1 type O strains, in a biological sample,said kit comprising at least one nucleic acid fragment as describedabove. This nucleic acid fragment may be used as a primer or a probe insaid kit.

Moreover, the present invention also provides for a kit for determiningthe presence of anti-HIV-1 type O) antibodies present in a biologicalsample, comprising at least one antigen or antigen fragment as describedabove.

In addition, the present invention provides for a kit for determiningthe presence of HIV-1 type O antigens present in a biological sample,comprising at least one antibody as described above.

The current invention also provides for a vaccine composition whichprovides protective immunity against HIV-1 infection, in particularagainst HIV-1 group O infection, comprising as an active principle atleast one antigen or antigen fragment as described above, or at leastone nucleic acid as described above, or a virus like particle (VLP)comprising at least one antigen or antigen fragment as described above,or an attenuated form of at least one of the HIV-1 type O strains asdescribed above, said active principle being combined with apharmaceutically acceptable carrier.

In a specific embodiment, polynucleic acid sequences coding for any ofthe antigens or antigen fragments as defined above, are used as avaccine, either as naked DNA or as part of recombinant vectors. In thiscase, it is the aim that said nucleic acids are expressed intoimmunogenic protein/peptide and thus confer in vivo protection to thevaccinated host (e.g. Ulmer et al., 1993).

The active ingredients of such a vaccine composition may be administeredorally, subcutaneously, conjunctivally, intramuscularly, intra nasally,or via any other route known in the art including for instance via thebinding to carriers, via incorporation into liposomes, by addingadjuvants known in the art, etc.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Amino acid alignment of gp41 sequences from the HIV-1 group Ostrains of the current invention, compared to gp41 sequences from someknown prototype HIV-1 group O strains (Ant70, MVP5180, VAU: boxed). Ifthe name of a strain is followed by -P or -PBMC, it means that thesequence was performed on strains present in peripheral blood monocytessamples, in stead of serum samples. Asteriks show perfectly conservedamino acids. Dots show well conserved amino acids. Dashes refer to gapsintroduced to maximize the alignment. The immunodominant domain isunderlined and within this domain the dashed line indicates theimmunosuppressive peptide (ISU) and the dotted line indicates theprincipal immunodominant domain (PID) by analogy to HIV-1 group Mviruses. The number of potential N-linked glycosylation sites which areshown by symbol {circumflex over ( )} above the amino acid alignment,are indicated at the right of the sequences on FIG. 1 (contd. 1).

FIG. 2. Amino acid alignment of C2V3 sequences originating from some ofthe HIV-1 group O strains of the current invention (189, FABA, MP340,MP450, MP448, MP539), compared to C2V3 sequences from some knownprototype HIV-1 group O strains (Ant70, MVP5180 and VAU: boxed).Asteriks show perfectly conserved amino acids. Dots show well conservedamino acids. Dashes refer to gaps introduced to maximize the alignment.The symbol + indicates the two conserved cysteine residues flanking theV3 loop region.

FIG. 3. Nucleic acid alignment of gp41 sequences originating from theHIV-1 group O strains of the current invention, compared to gp41sequences from some known prototype HIV-1 group O strains (Ant70,MVP5180, VAU and VI686: in bold). Asteriks show positions of conservednucleic acids. Dashes refer to gaps introduced to maximize alignment.

FIG. 4. Nucleic acid alignment of C2V3 sequences originating from someof the HIV-1 group O strains of the current invention (189, FABA, MP340,MP448, MP450 and MP539), compared to C2V3 sequences from some knownprototype HIV-1 group O strains (MVP5 180, Ant70 and V1686: in bold).Asteriks show positions of conserved nucleic acids. Dashes refer to gapsintroduced to maximize alignment.

FIG. 5. Phylogenetic tree analysis for the gp41-sequenced region of thenew HIV-1 group O strains of the current invention, compared to theprototype HIV-1 group O strains (Ant70, MVP5180, VAU and VI686), HIV-1group M strains (U455, Z2Z6 and MN), and SIVcpz-strains. SIVcpz-ANT hasbeen used as an “outgroup” for the analysis, and is therefore putbetween brackets [ ].The viruses of the current invention are indicatedin bold. Country of origin is mentioned between parentheses.Phylogenetic relationships were determined using the neighbor joiningmethod, as described in Materials and Methods. The numbers given at thebranch points represent bootstrap values out of 100 obtained for theneighbor joining method.

FIG. 6 Nucleotide and amino acid sequences obtained from the new HIV-1group O strains of the current invention.

FIG. 7.

7A. Comparison of the consensus amino acid sequences of the potentialgp41-immunosuppressive peptide (ISU) for HIV-1 group M and O strains andISU-peptide for SIVcpzGAB and SIVcpzANT.

7B. Antigenicity/hydrophilicity plots of the consensus ISU-peptide (17amino acids flanked by Leucine (L) residues) for HIV-1 group O and groupM viruses. A value of 100% or nearly predicts the considered peptide tobe highly immunogenic.

FIG. 8.

8A. Nucleic acid sequence (SEQ ID NO 106) and the corresponding aminoacid sequence translation of part of the genome of HIV-1 group O virusMP645. The corresponding polypeptides Pol (partially) (SEQ ID NO130, Vif(SEQ ID NO 110), Vpr (SEQ ID NO 118), Tat (SEQ ID NO 122), Rev (SEQ IDNO 126), Vpu (SEQ ID NO 114) and Env (SEQ ID NO 134) (partially) areunderlined and their corresponding name is indicated at the right ofeach open reading frame.

8B. Nucleic acid sequence (SEQ ID NO 103) and the corresponding aminoacid sequence translation of part of the genome of HIV-1 group O virusMP331 (FABA). The corresponding polypeptides Pol (partially) (SEQ ID NO127), Vif (SEQ ID NO 107), Vpr (SEQ ID NO 115), Tat (SEQ ID NO 119), Rev(SEQ ID NO 123), Vpu (SEQ ID NO 111) and Env (partially) (SEQ ID NO 131)are underlined and their corresponding name is indicated at the right ofeach open reading frame.

8C. Nucleic acid sequence (SEQ ID NO 104) and the corresponding aminoacid sequence translation of part of the genome of HIV-1 group O virusMP448. The corresponding polypeptides Pol (partially) (SEQ ID NO 128),Vif (SEQ ID NO 108), Vpr (SEQ ID NO 116), Tat (SEQ ID NO 120), Rev (SEQID NO 124), Vpu (SEQ ID NO 112) and Env (partially) (SEQ ID NO 132) areunderlined and their corresponding name is indicated at the right ofeach open reading frame.

8D. Nucleic acid sequence (SEQ ID NO 105) and the corresponding aminoacid sequence translation of part of the genome of HIV-1 group O virusMP539. The corresponding polypeptides Pol (partially) (SEQ ID NO 129),Vif (SEQ ID NO 109), Vpr (SEQ ID NO 117), Tat (SEQ ID NO 121), Rev (SEQID NO 125), Vpu (SEQ ID NO 113) and Env (partially) (SEQ ID NO 133) areunderlined and their corresponding name is indicated at the right ofeach open reading frame.

TABLE LEGEND

Table 1:

Divergence between HIV-1 group M and O viruses and the chimpanzeeviruses SIVcpzGAB and SIVcpzANT based on gp41 nucleic acid sequences.

^(a): Divergence from group M viruses was calculated for at least threerandomly selected strains for each of the subtypes from A to G.

Table 2:

Percent divergence (=100%-% homology) between the gp41 nucleic acidsequences of the different HIV-1 group O strains of the currentinvention as specified in FIG. 3.

TABLE 1 % genetic divergence Group O Group M CPZANT CPZGAB 37.5(35.2-38.8) 31.2 (29.3-32.5) 32.6 CPZANT 36.5 (32.4-39.1) 33.7(32.9-34.3) Group M 37.3 (35.0-40.8) 12.3 (2.2-16.6) Group O 14.7(1.2-21.8)

TABLE 2 MP340 FABA MP450 MP448 ANT70 MVP5180 VAU V1686 (PBMC) MP340(PBMC) FABA (PBMC) MP450 (PBMC) MP448 189 ANT70 — 18 16  9 13 13 12 1312 12 10  9 14 MVP5180 — 17 22 24 23 24 25 23 23 16 15 26 VAU — 18 21 2120 21 20 20 18 17 22 V1686 — 14 14 15 16 14 14 10 10 16 MP340 (PBMC) — 1 14 15  2  2 14 14  2 MP340 — 13 12  2  2 15 15  2 FABA (PBMC) —  5 1313 15 15 13 FABA — 13 13 15 15 15 MP450 (PBMC) —  0 13 13  3 MP450 — 1313  3 MP448 (PBMC) —  0 16 MP448 — 16 189 — 320 BSD422 KGT008 MP575BSD189 BSD649 BSD242 533 772P94 MP95B MP539 320 BSD422 KGT008 MP575BSD189 BSD649 BSD242 533 772P94 MP95B MP539 ANT70  9  9 11  7  9  6 2223 19 15 18 MVP5180 21 21 23 21 21 20  7  8 15 13 22 VAU 18 18 18 16 1715 18 18 15 16 15 V1686 11 12  9 10 10 10 26 25 22 15 18 MP340 (PBMC) 1415 17 13 11 12 26 24 20 20 20 MP340 14 14 17 13 11 12 27 24 20 20 19FABA (PBMC) 14 18 17 13 14 13 28 26 20 20 22 FABA 15 18 17 14 14 14 2927 20 20 22 MP450 (PBMC) 12 15 17 12 10 12 26 24 19 19 20 MP450 12 15 1712 10 12 26 24 19 19 20 MP448 (PBMC) 10 12 12 10 11 11 19 19 19 15 17MP448 11 12 12  9 11 11 18 19 18 14 17 189 15 17 19 14 13 14 28 26 22 2222 320 — 12 11  7 11  8 22 22 22 16 18 BSD422 — 15 11 13 10 24 26 19 1518 KGT008 — 10 12 12 26 26 22 19 20 MP575 —  8  7 25 25 20 17 17 BSD189—  9 26 26 19 17 18 BSD649 — 24 23 19 16 18 BSD242 —  5 16 16 25 533 —18 16 23 772P94 — 17 19 MP95B — 18 MP539 —

EXAMPLES Example 1 Materials and Methods

Patients and Viruses

A total of 16 viruses have been characterized. Patients were identifiedas being infected with an HIV-1 group O virus using a specificserological testing algorithm, based on V3 peptides from different M andO strains (consensusM, M-Mal, O-ANT-70, O-VI686, O-MVP5180 (INNOLIAHIV-1 type O Research product, Innogenetics, Belgium), as describedelsewhere (Peeters et al., in press). Ten patients were from Cameroon(BSD189, BSD242, BSD422, BSD649, MP340, MP95B, MP448, MP575, MP539 andMP450), 2 from Gabon (189, 533) and the others from Tchaad (320),Nigeria (KGT008), Senegal (FABA=331) and Niger (772P94). For 3 patientsfrom Cameroon (MP340, MP448 and MP450) and for the patients from Nigeriaand Senegal primary uncultured peripheral blood mononuclear cells(PBMCs) were available, while for the other patients from Cameroon,Gabon, Niger and Tchaad only serum was available. Strain V1686 fromGabon has been described previously (Janssens et al. 1994).

Nucleic Acid Extraction

DNA was extracted from PBMCs using the IsoQuick isolation kit(Microprobe Corp., Garden Cove, Calif., USA), resuspended in theappropriate volume of nuclease free water and 1/10 was used foramplifications. Viral RNA was extracted from 50 μl of plasma by theguanidinium thiocyanate-phenol-chloroform method as described previouslyby Chomczynsky and Sacchi (1987), resuspended in 5 μl of nuclease freewater and further used for reverse transcription reaction.

RT, PCR and Sequencing

The reverse transcription reaction (RT) was performed in a final volumeof 20 μl, containing 50 mM TrisHCl pH 8.3, 50 mM KCl, 10 mM MgCl₂, 10 mMDTT, 0.5 mM spermidine, 1 mM each deoxynucleoside triphophate, 0.5 μMouter reverse primer (41-4, see further) and 5 U of Avian MyeloblastosisVirus Reverse Transcriptase (Promega), for 30 min at 42° C. Fivemicroliters from the RT reaction were used for PCR amplification.

Nested PCR was used to amplify a fragment of approximately 420 bp fromthe gp41-region. Outer primers allow amplification of HIV-1 sequencesfrom group O and M (sense 41-1: 5′-GGGTTCTTGGGAGCAGCAGGAAGCACTATGGGCG-3′(SEQ ID NO 139), antisense 41-4: 5′-TCTGAAACGACAGAGGTGAGTATCCCTGCCTAA-3′(SEQ ID NO 140)). Inner primers were determined according to theHIV-1-Ant70 sequence (Vanden Hae-sevelde et al. 1994) (sense 41-6:5′-TGGATCCCACAGTGTACTGAAGGGTATAGTGCA-3′ (SEQ ID NO 141), antisense 41-7:5′-CATTTAGTTATGTCAAGCCAATTCCAAA-3′ (SEQ ID NO 142)). PCRs were performedin a final volume of 100 μl containing 10 mM Tris-HCl (pH 9.0), 50 mMKCl, 1.5 mM MgCl2, 0.2 mM each deoxynucleoside triphosphate, 2.5 U ofTaq DNA polymerase (Promega) and 0.4 μM of each primer. After an initialdenaturation step of 3 min at 94° C., 30 to 35 cycles were performed at94° C. for 1 min to 20 s, 50° C. for 1 min to 30 s, 72° C. for 1 min,followed by a final extension of 10 to 7 min. For the second round, 1 to5 μl of the first amplification were subjected to the same cyclingconditions for 35 to 40 PCR cycles.

Amplification of the C2V3-region was obtained by nested PCR using a setof primers selected from the HIV-1-Ant70 sequence. Outer primers were:anti-sense V70-5 (5′-GTTCTCCATATATCTTTCATATCTCCCCCTA-3′, SEQ ID NO 143)and sense V70-1 (5′-TTGTACACATGGCATTAGGCCAACAGTAAGT-3′, SEQ ID NO 144)and inner primers were: sense V70-2(5′-TGAATTCCTAATATTGAATGGGACACTCTCT-3′, SEQ ID NO 145) and antisenseV70-4 (5′-TGGATCCTACAATAAAAGAATTCTCCATGACA-3′, SEQ ID NO 146).Amplification conditions were as described above.

Each fragment was sequenced on both strands, as previously described(Bibollet-Ruche 1997), using an Applied Biosystems sequencer (model373A, Applied Biosystems, Inc) and a dye-deoxy terminator procedure, asspecified by the manufacturer.

Sequence Analysis

Overlapping sequences were joined by using SeqEd-1.0 (AppliedBiosystems, Inc). Sequences were aligned using CLUSTAL V (Higgins et al1992; Higgins and Sharp 1988) program. Evolutionary distances werecalculated by using the Kimura's two-parameters method with correctionfor the multiple substitutions and excluding positions with gaps inaligned sequences (Kimura 1983). Phylogenetic relationships werecomputed from the distance matrix by the neighbor-joining method (Saitouand Nei 1987). Phylogenetic analyses were also performed by a parsimonyapproach and implemented using DNAPARS. In both cases, reliability ofthe branching orders was confirmed by the bootstrap approach(Felsenstein 1985). Phylogenetic analyses were also performed forproteic sequences using PROTPARS. These methods were implemented usingthe PHYLIP 3.56 package (Felsenstein 1993). The results were similarwith both methods, for nucleotidic and proteic sequences, in allessential aspects.

Antigenicity Profiles

Antigenicity of the ISU peptides have been calculated according toprograms developed by Garnier et al (1978) and Gibrat et al (1987).

Example 2 Nucleic Acid Sequences and Phylogenetic Analysis

Analysis of gp41 Sequences

For the 16 viruses, 330 to 351 bp of the region spanning theimmunodominant domain of the transmembrane gp41 glycoprotein (by analogyto HIV-1 group M viruses) was characterized. The different sequencesobtained are represented in FIG. 6. These sequences have been aligned tothe corresponding gp41-sequences of known HIV-1 group O strains, asshown in FIG. 3. Moreover, sequence identity was calculated for intraand intergroup similarity by pairwise alignment and comparison (Table1). For the group M viruses, these values were calculated for randomlyselected strains for which sequences were available in the database andwhich were representative of subtypes A to G (the same sequences werefurther used for phylogenetic analyses). Divergence between SIVcpz-GAB(Huet et al. 1990) and the group M (mean 31,2%) was lower thandivergence between SIVcpz-GAB and the group O (mean 37%). These resultswere similar when calculated with the second chimpanzee virus,SIVcpz-ANT (Vanden Haesevelde et al. 1996), with mean divergence of33.7% and 36.5% with group M and O respectively (Table 1). Intragroupdivergence for group O viruses ranged from 1.2 to 21.8% (mean 14.7%)while divergence within group M clades (A to G) ranged from 2.2 to 16.6%(mean 12.3%).

Phylogenetic trees were constructed with the 16 new sequences describedhere, the four group O strains of which the gp41 sequences have alreadybeen characterized, nl. Ant70 (Vanden Haesevelde et al. 1994), MVP5180(Gürtler et al. 1994), VAU (Charneau et al. 1994) and VI686 (Janssens etal. 1994) and group M viruses representative for A (U455), B (MN) and D(Z2Z6) subtypes (see FIG. 5). Trees were constructed from nucleotidicsequences according to the alignment in FIG. 3, by the neighbor-joiningand by the maximum-parsimony methods. Consistent results were obtainedby the two methods: group M and group O viruses clustered separately(bootstrap values 98 and 100 respectively).

Within the newly characterized HIV-1 type O strains new subclusters mayappear, such as for example the subcluster consisting of strains MP340,189 and MP450, which segregate with a bootstrap value of 100, or thesubcluster consisting of strains MVP5180, 533, BSD242 (bootstrap value98). These subclusters may define different genotypes within group O inanalogy to the different group M genotypes (clades). Phylogeneticanalyses were also performed for the deduced amino acid sequences forgroup O and M viruses (data not shown) and the results were consistentwith the nucleotidic sequences, although lower bootstrap values wereobserved (approximately 65%), probably due to the short size of thesequences (109 a.a., gaps excluded, data not shown). The countries fromwhich these viruses were sampled are indicated alongside the strains andno links were found between the manner of clustering and thegeographical origins of the strains.

Table 2 shows the percentage divergence between gp41-nucleotidesequences originating from HIV-1 group O strains of the invention andknown HIV-1 group O strains (Ant70, MVP5180, VAU and VI686).

Analysis of C2V3 Sequences

For 6 of the 16 new virus strains (189, FABA, MP340, MP448, MP450 andMP539), C2V3 sequences were determined. FIG. 4 shows the alignment ofthese new C2V3 sequences with the corresponding sequences of known HIV-1group O strains.

Example 3 Analysis of Deduced Amino Acid Sequences

gp41 Amino Acid Sequences

FIG. 1 depicts the alignment of the deduced gp41 amino acid sequences.Although different from group M, this region of the transmembraneglycoprotein is also highly conserved among group O viruses, especiallyfor the region corresponding to the immunodominant region in group Misolates. This region has been divided into two domains, correspondingto an immunosuppressive peptide (ISU-peptide) of 17 amino acids (aa) andto an immunodominant cysteine loop of 7 aa (Oldstone et al. 1991), alsocalled the principal immunodominant domain (PID). The consensus cysteineloop for group O viruses (CKGR/KLV/IC) is quite closely related to thegroup M consensus (CSGKLIC), suggesting that they could have similarfunctions in both virus groups. This PID sequence, corresponding to aB-cell epitope recognized by nearly all sera from patients or animals(Bertoni et al. 1994; Chong et al. 1991; Gnann et al. 1987; Norrby etal. 1987; Pancino et al. 1993), seems to be a very conserved structure,in contrast to the variability observed for the neutralizing epitope ofthe V3 loop of HIV-1 gp120 (Zwart et al. 1991; Zwart et al. 1993).Deletion of this PID peptide or disruption of the cysteine loop severelyimpairs the gp160 processing, leading to a loss of infectivity in HIV-1(Dedera et al. 1992, Pancino et al. 1993; Schulz et al. 1992) and alsoin the far related type D simian retrovirus (Brody and Hunter 1992).

Comparison of ISIU-peptides from the group M, SIVcpz and group O virusesare presented in FIG. 7A. The consensus peptide from group O is quitedivergent from the SIVcpz/group M peptide by the presence of an arginine(R, positively charged) in position 2 instead of a phenylalanine (Q,hydrophilic), a leucine (L) in position 5 and 8 instead of a valine (V),and a very different stretch TLIQN instead of RYLKD in position 10-14.Prediction of the secondary structure revealed an alpha helix in eachcase, and the predicted isoelectric point is 9,7 and 11,3 for groupM/SIVcpz and group O strains respectively. FIG. 7B represents thepredicted hydrophilicity/antigenicity plot and revealed the presence ofa second peak at position 3 to 7 (ARLLA) for group O in addition to thepeak at position 12 (L) conserved in group M/SIVcpz and group O viruses.

The divergence of the ISU peptide between group O and M viruses maysuggest different functionalities of the ISU peptide in both groups. Forseveral retroviruses this ISU peptide has been shown to suppress a broadrange of immune reactions such as the inhibition of IL-2-dependant andconcanavaline A-induced proliferation of T lymphocytes (Denner et al.1994; Ruegg et al. 1989) but also B lymphocyte proliferation for HIV-1group M (Denner et al. 1996). The principal B-epitope of this ISUpeptide has been mapped at the C-terminal end, centered around theleucine residue in position 12 for group M (Denner et al. 1994). Thedata of FIG. 7 show a similar antigenicity peak for group O viruses but,in contrast to group M and SIVcpz, the presence of a second peak isobserved towards the N-terminal part of this peptide. Moreover, antibodyresponse against this region in HIV-1 group M infected individuals hasbeen shown to be vigorous and independent of the stage of the disease(Cumming et al. 1990; Zwart et al. 1994). This result raises thepossibility of the design of peptides for the specific detection ofantibodies against group O viruses, which could improve thediscrimination between the two HIV-1 groups, and could also be usefulfor the detection of divergent group O viruses given the highconservation of these peptides among the different strains we havecharacterized.

A third interesting feature concerns the number and the position of theN-linked glycosylation sites in the immunodominant domain (FIG. 1). Ingroup M viruses, 4 sites are conserved in the gp41 extracellular domainand they have been shown to play important roles in the intracellularprocessing of the gp160 and in the fusogenic properties of gp41 (Dederaet al. 1992; Vanini et al. 1993). Most of the group O viruses describedhere contain only three potential N-linked glycosylation sites whereasonly a few strains contain four. Only the position of the first site ishighly conserved (position 60 in the alignment) and the positions of thetwo or three other sites vary among the strains, in contrast to group Mviruses.

In HIV-1 group M viruses, it is known that removal of the thirthglycosylation site abolishes the cleavage of the gp160 precursor andtransport to the plasma membrane of infected cells. An immediateconsequence of this result, obtained in vitro after transfection of thismutant in HeLa/CD4 cells, is that it fails to induce syncytia (Dash etal. 1994). Roles of the other N-linked glycosylation sites of the gp41have not been filly explored but their high conservation suggestsimportant functions. In the case of the gp120, the highly conservedsites surrounding the V3 loop have been shown to influence infectivity(Lee et al. 1992), protein folding (Li et al. 1993) and immunogenicity(Benjouad et al 1992) of this domain. For group O viruses, only thefirst site is conserved in all the strains which were characterized.Even if the importance of N-glycosylation has not been studied for HIV-1group O viruses, it can be speculated that differences will be observedwhen compared to group M viruses.

In conclusion, comparison of the gp41 amino acid sequences clearlydiscriminate the two HIV-1 groups, either by the pattern of N-linkedglycosylation sites or by the characteristics of the ISU peptide of theectodomain of the transmembrane glycoprotein. The differences observedbetween the two HIV-1 groups in the gp41 region raise the possibilitythat these viruses use different mechanisms, for example in theintracellular processing of the envelope precursor or in the domainsinvolved in the membrane fusion. Several studies have suggested that theimmunodominant region of the gp41 in group M viruses, which is a targetfor neutralizing but also enhancing antibodies, could be a promisingcandidate for vaccine development (Cumming et al. 1990; Muster et al.1993; Zwart et al. 1994). The data disclosed in the current inventionsuggest that a vaccine developed against HIV-1 group M viruses could beineffective against group O strains, especially when these viruses coulduse different pathways to induce immunodeficiency in infected patients.

Analysis of C2V3 Amino Acid Sequences

FIG. 2 depicts the alignment of the deduced C2V3 sequences of 6 of theHIV-1 group O strains of the invention as compared to known HIV-1 groupO strains. The delineation of the C2 and V3 region is according toStarcich et al. (1986) and Willey et al. (1986). The C2 region belongsto the rather conserved regions which are important in protein foldingand protein function. The V3 region is one of the hypervariable regions,for which amino acid conservation between different HIV-1 isolates isbelow 50%. Hypervariable regions also contain short deletions orinsertions (Myers et al. 1992).

The V3-region of gp120, also known as the principal neutralizingdeterminant (PND), contains a loop of 35 amino acids, formed by acysteine-cysteine disulfide bridge. The PND is implicated in severalimportant biological functions, such as:

1. it determines the HIV-1 cell tropism (Hwang et al. 1991, Shioda etal. 1991)

2. it affects fusion (Feed et al. 1991) and viral virulence (Fouchier etal. 1992)

3. it stimulates cytotoxicity (Takahashi et al. 1992)

4. it is the primary target for antibody neutralization (Palker et al.1989).

The fact that the V3 domain of gp120 may induce a protective immuneresponse, has made this region of particular interest for vaccineresearch. Although the V3 region as a whole is hypervariable, there is arather high sequence conservation in its tetrapeptide sequenceGly-Pro-Gly-Arg (GPGR) at the crown of the loop (LaRosa et al. 1990).This motif corresponds to the binding site of the neutralizingantibodies, and single amino acid changes within this epitope highlyreduce antibody binding (Meloen et al. 1989).

The alignment in FIG. 2 shows that sequence variability within the V3loop is high between the different HIV-1 group O strains, includingthose of the current invention. These new sequences, and moreparticularly the sequences represented by SEQ ID NO 53 to 70 and SEQ IDNO 83 to 90 are important in the design of new and better (=moresensitive and/or more specific) HIV-diagnostic assays. Moreover, thevarious sequences in the V3 region will be important with regard to thedevelopment of HIV-vaccines providing an as broad as possible protectionagainst HIV-infection.

Example 4 Cloning Strategy to Obtain Vif, Vpu, Vpr, Tat, Rev, Env andPol

Polymerase Chain Reaction Amplification and Sequencing

The sequences of the nested primer sets were designed on HIV-1 nucleicacid sequences in conserved regions flanking the Vif and Vpu genes. DNAfrom cultured and uncultured PBMCs was extracted using IsoQuick(Microprobe, Garden Cove, Calif.) according to the manufacturerinstructions and quantified spectrophotometrically. Approximately 1 μgof DNA was used for a first round of amplification with an outer primerpair (VIF1, 5′ GGGTTTATTACAGGGACAGCAGAG 3′ (SEQ ID NO 147) and VPU1, 5′GGTTGGGGTCTGTGGGTACACAGG 3′) (SEQ ID NO 148) in a final volume of 100 μlof containing 10 mM Tris-HCl (pH 9.0), 50 mM KCl, 1.5 mM MgCl₂ a 0.2 mMconcentration of each deoxynucleoside triphosphate, 2.5 U of Taq DNApolymerase (Promega, Madison, Wis.), and a 0.4 μM concentration of eachprimer. Five microliters from this first round was used for a secondround with an inner primer pair (VIF2, 5′ GCAAAACTACTCTGGAAAGGTG 3′ (SEQID NO 149) and VPU2, 5′ GCWTCTTTCCACACAGGTACCCC 3′ (SEQ ID NO 150) whereW represents an A or a T) under the same reaction conditions. The tworounds of PCR were run for 35 cycles each under the following cyclingconditions:94° C. for 30 sec, 50° C. for 30 sec, and 72° C. for 2 mn.The two rounds of PCR were preceded by a denaturation step of 3 min at94° C. and followed by a final extension step of 7 min at 72° C.

Sequencing of the amplified products was done directly afterpurification by TAE-low melting point agarose gel electrophoresis(Bibollet-Ruche et al, 1997) using an Applied Biosystems (Foster City,Calif.) 373 Stretch sequencer and a Dye-Deoxy terminator procedure (dyeterminator cycle sequencing ready reaction, with AmpliTaq DNApolymerase; Perkin-Elmer. Norwalk, Conn.) as specified by themanufacturer. Inner polymerase chain reaction (PCR) primers (VIF2 andVPU2) and inner sequencing primers (OVIF, 5′ CATATTGGGGATTGATGCCAG 3′(SEQ ID NO 151); OVPU, 5′ GCATYAGCGTTACTTACTGC 3′: Y=C or T (SEQ ID NO152)) were used. Overlapping sequences were joined using SeqEd (AppliedBiosystems) to obtain the full-length sequence. Direct sequencing wasperformed on PCR-generated fragments. Ambiguities observed at a limitednumber of positions in some sequences were resolved when joining theoverlapping fragments.

Analyses of Accessory Protein Sequences

Open reading frames for the different accessory proteins (Pol, Vif, Vpr,the first exon of Tat, Vpu and Env) were determined and the deducedprotein sequences were obtained using the Translate program option ofthe PCgene software package. The resulting sequences are indicated inFIG. 8 (B to D). FIG. 8A shows the sequences of HIV-1 group O strainMP645 which was obtained following a similar approach as the onedescribed above for MP448, MP539 and MP331.

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SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 152 <210> SEQ ID NO 1 <211>LENGTH: 340 <212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 1aacctgctaa gagcaataca ggcccagcaa gaattgctga ggctatctgt atggggtatc 60agacaactcc gagctcgcct gctagcctta gaaaccttaa tacagaatca gcagctccta 120aacctatggg gttgtaaggg aaggatagtc tgctacacat cagtaaaatg gaacgataca 180tggagacatg tcactaatat gagtgaagtt tgggacaaac taacctggca ggaatgggat 240cggcagatag acaacataag ctatgttata tatgatgaaa tacaaagagc acaagtacag 300caagaacaaa atgagaagaa gttgctggag ttagatgaat 340 <210> SEQ ID NO 2 <211>LENGTH: 113 <212> TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE: 2 AsnLeu Leu Arg Ala Ile Gln Ala Gln Gln Glu Leu Leu Arg Leu Ser 1 5 10 15Val Trp Gly Ile Arg Gln Leu Arg Ala Arg Leu Leu Ala Leu Glu Thr 20 25 30Leu Ile Gln Asn Gln Gln Leu Leu Asn Leu Trp Gly Cys Lys Gly Arg 35 40 45Ile Val Cys Tyr Thr Ser Val Lys Trp Asn Asp Thr Trp Arg His Val 50 55 60Thr Asn Met Ser Glu Val Trp Asp Lys Leu Thr Trp Gln Glu Trp Asp 65 70 7580 Arg Gln Ile Asp Asn Ile Ser Tyr Val Ile Tyr Asp Glu Ile Gln Arg 85 9095 Ala Gln Val Gln Gln Glu Gln Asn Glu Lys Lys Leu Leu Glu Leu Asp 100105 110 Glu <210> SEQ ID NO 3 <211> LENGTH: 340 <212> TYPE: DNA <213>ORGANISM: Human <220> FEATURE: <221> NAME/KEY: misc_feature <222>LOCATION: (1)..(340) <223> OTHER INFORMATION: N = any nucleotide <400>SEQUENCE: 3 aacctgctaa sancaataca ggcccakcaa gaattgctga ggctatctgtatggggtatc 60 agacaamtcc gagstygcct gstagcctta gaaaccttaa tacasaatcagcasctccta 120 aacctatggg gttgtaaagg aaggatastn tgctacacat cagtaaaatggaacnataca 180 tggaaacatg tcactnatat gagtgaagtt tgggacaaac taacctggcaggaatgggat 240 cggcngatag acaacataag ctatgttata tatgatgnna tacaaagagcacaagtacag 300 caagaacaaa atgagaagaa gttgctggag ttagatgaat 340 <210> SEQID NO 4 <211> LENGTH: 113 <212> TYPE: PRT <213> ORGANISM: Human <220>FEATURE: <221> NAME/KEY: Misc_feature <222> LOCATION: (1)..(113) <223>OTHER INFORMATION: Xaa = unknown <400> SEQUENCE: 4 Asn Leu Leu Xaa XaaIle Gln Ala Xaa Gln Glu Leu Leu Arg Leu Ser 1 5 10 15 Val Trp Gly IleArg Gln Xaa Arg Xaa Xaa Leu Xaa Ala Leu Glu Thr 20 25 30 Leu Ile Xaa AsnGln Xaa Leu Leu Asn Leu Trp Gly Cys Lys Gly Arg 35 40 45 Ile Xaa Cys TyrThr Ser Val Lys Trp Asn Xaa Thr Trp Lys His Val 50 55 60 Thr Xaa Met SerGlu Val Trp Asp Lys Leu Thr Trp Gln Glu Trp Asp 65 70 75 80 Arg Xaa IleAsp Asn Ile Ser Tyr Val Ile Tyr Asp Xaa Ile Gln Arg 85 90 95 Ala Gln ValGln Gln Glu Gln Asn Glu Lys Lys Leu Leu Glu Leu Asp 100 105 110 Glu<210> SEQ ID NO 5 <211> LENGTH: 352 <212> TYPE: DNA <213> ORGANISM:Human <400> SEQUENCE: 5 aacctgctaa aagcaataca ggcccagcag caattgctgaggttatctgt atggggtatc 60 aaacaactcc gagctcgcct gctagcctta gaaaccttaatacagaatca gcaactccta 120 aacctatggg gctgtaaagg aaggctagtc tgctacacatcagtaaaatg gaacaataca 180 tggacaaaaa acatcacaaa catcacagac ctagacgagatttgggacaa atttacatgg 240 caccaatggg atcaacagat aaaccacata agtgatgtcatatatgaaga aataccaaag 300 gcacaagtac agcagggacc aaatgagagg aagttgctggagttagatga at 352 <210> SEQ ID NO 6 <211> LENGTH: 117 <212> TYPE: PRT<213> ORGANISM: Human <400> SEQUENCE: 6 Asn Leu Leu Lys Ala Ile Gln AlaGln Gln Gln Leu Leu Arg Leu Ser 1 5 10 15 Val Trp Gly Ile Lys Gln LeuArg Ala Arg Leu Leu Ala Leu Glu Thr 20 25 30 Leu Ile Gln Asn Gln Gln LeuLeu Asn Leu Trp Gly Cys Lys Gly Arg 35 40 45 Leu Val Cys Tyr Thr Ser ValLys Trp Asn Asn Thr Trp Thr Lys Asn 50 55 60 Ile Thr Asn Ile Thr Asp LeuAsp Glu Ile Trp Asp Lys Phe Thr Trp 65 70 75 80 His Gln Trp Asp Gln GlnIle Asn His Ile Ser Asp Val Ile Tyr Glu 85 90 95 Glu Ile Pro Lys Ala GlnVal Gln Gln Gly Pro Asn Glu Arg Lys Leu 100 105 110 Leu Glu Leu Asp Glu115 <210> SEQ ID NO 7 <211> LENGTH: 352 <212> TYPE: DNA <213> ORGANISM:Human <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION:(1)..(352) <223> OTHER INFORMATION: N = any nucleotide <400> SEQUENCE: 7nacctgttaa gagcaataca ggcccagcag caattggtga ggttatctgt atggggtatc 60agacaaatcc gaggtngcct ggtagcctta gaaaccttaa tacagaatca gcaantcctn 120aacctatggg gctgtaaagg aagggtagtt tgntacacat cagtaaaatg gaacaataca 180tggacaaaaa acatcacaaa catcacagac ctagacgaga tttgggacaa atttacatgg 240cagcaatggg atcaacagat aaacaacata agtgatgtcc tatatgaaga aatacaaaag 300gcacaagtac agcaggaaca aaatgagagg aagttgctgg agttagatga at 352 <210> SEQID NO 8 <211> LENGTH: 117 <212> TYPE: PRT <213> ORGANISM: Human <220>FEATURE: <221> NAME/KEY: Misc_feature <222> LOCATION: (1)..(117) <223>OTHER INFORMATION: Xaa = Unknown <400> SEQUENCE: 8 Xaa Leu Leu Arg AlaIle Gln Ala Gln Gln Gln Leu Val Arg Leu Ser 1 5 10 15 Val Trp Gly IleArg Gln Ile Arg Gly Xaa Leu Val Ala Leu Glu Thr 20 25 30 Leu Ile Gln AsnGln Gln Xaa Xaa Asn Leu Trp Gly Cys Lys Gly Arg 35 40 45 Val Val Xaa TyrThr Ser Val Lys Trp Asn Asn Thr Trp Thr Lys Asn 50 55 60 Ile Thr Asn IleThr Asp Leu Asp Glu Ile Trp Asp Lys Phe Thr Trp 65 70 75 80 Gln Gln TrpAsp Gln Gln Ile Asn Asn Ile Ser Asp Val Leu Tyr Glu 85 90 95 Glu Ile GlnLys Ala Gln Val Gln Gln Glu Gln Asn Glu Arg Lys Leu 100 105 110 Leu GluLeu Asp Glu 115 <210> SEQ ID NO 9 <211> LENGTH: 340 <212> TYPE: DNA<213> ORGANISM: Human <400> SEQUENCE: 9 aacctgctaa gagcaataca ggcccagcagcaattgctga ggctatctgt atggggtatc 60 agacaactcc gagctcgcct gctagccttagaaaccttaa tacagaatca gcagctccta 120 aacctatggg gttgtaaggg aaggatagtctgctacacat cagtaaaatg gaacaataca 180 tggagaaatg tcactaatat gagtgaagtttgggacacac taacctggca ggaatgggat 240 cggcagatag acaacataag ctatgttatatatgatgaaa tacaaagagc acaagtacag 300 caggaacaaa atgagaagaa gttgctggagttagatgaat 340 <210> SEQ ID NO 10 <211> LENGTH: 113 <212> TYPE: PRT<213> ORGANISM: Human <400> SEQUENCE: 10 Asn Leu Leu Arg Ala Ile Gln AlaGln Gln Gln Leu Leu Arg Leu Ser 1 5 10 15 Val Trp Gly Ile Arg Gln LeuArg Ala Arg Leu Leu Ala Leu Glu Thr 20 25 30 Leu Ile Gln Asn Gln Gln LeuLeu Asn Leu Trp Gly Cys Lys Gly Arg 35 40 45 Ile Val Cys Tyr Thr Ser ValLys Trp Asn Asn Thr Trp Arg Asn Val 50 55 60 Thr Asn Met Ser Glu Val TrpAsp Thr Leu Thr Trp Gln Glu Trp Asp 65 70 75 80 Arg Gln Ile Asp Asn IleSer Tyr Val Ile Tyr Asp Glu Ile Gln Arg 85 90 95 Ala Gln Val Gln Gln GluGln Asn Glu Lys Lys Leu Leu Glu Leu Asp 100 105 110 Glu <210> SEQ ID NO11 <211> LENGTH: 340 <212> TYPE: DNA <213> ORGANISM: Human <400>SEQUENCE: 11 aacctgctaa gagcaataca ggcccagcag caattgctga ggctatctgtatggggtatc 60 agacaactcc gagctcgcct gctagcctta gaaaccttaa tacagaatcagcagctccta 120 aacctatggg gttgtaaggg aaggatagtc tgctacacat cagtaaaatggaacaataca 180 tggagaaatg tcactaatat gagtgaagtt tgggacacac taacctggcaggaatgggat 240 cggcagatag acaacataag ctatgttata tatgatgaaa tacaaagagcacaagtacag 300 caggaacaaa atgagaagaa gttgctggag ttagatgaat 340 <210> SEQID NO 12 <211> LENGTH: 113 <212> TYPE: PRT <213> ORGANISM: Human <400>SEQUENCE: 12 Asn Leu Leu Arg Ala Ile Gln Ala Gln Gln Gln Leu Leu Arg LeuSer 1 5 10 15 Val Trp Gly Ile Arg Gln Leu Arg Ala Arg Leu Leu Ala LeuGlu Thr 20 25 30 Leu Ile Gln Asn Gln Gln Leu Leu Asn Leu Trp Gly Cys LysGly Arg 35 40 45 Ile Val Cys Tyr Thr Ser Val Lys Trp Asn Asn Thr Trp ArgAsn Val 50 55 60 Thr Asn Met Ser Glu Val Trp Asp Thr Leu Thr Trp Gln GluTrp Asp 65 70 75 80 Arg Gln Ile Asp Asn Ile Ser Tyr Val Ile Tyr Asp GluIle Gln Arg 85 90 95 Ala Gln Val Gln Gln Glu Gln Asn Glu Lys Lys Leu LeuGlu Leu Asp 100 105 110 Glu <210> SEQ ID NO 13 <211> LENGTH: 331 <212>TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 13 aacctgctaa gagcaatacaggcccagcag cacttgctga ggctatctgt atggggtatc 60 agacaactcc gagctcgcctgctagcctta gaaaccttaa tacagaatca gcaactccta 120 aactcatggg gctgtaagggaaagatagtc tgttacacag cagtaaaatg gaacaagaca 180 tggacaggaa atgaaagtatttgggaccac ctcacatggc agcaatggga tcagcagata 240 gacaatgtaa gctccaccatatatgaggaa atactaaaag cacaagtaca gcaggaacag 300 aatgagcaaa agttgctggagttagatgaa t 331 <210> SEQ ID NO 14 <211> LENGTH: 110 <212> TYPE: PRT<213> ORGANISM: Human <400> SEQUENCE: 14 Asn Leu Leu Arg Ala Ile Gln AlaGln Gln His Leu Leu Arg Leu Ser 1 5 10 15 Val Trp Gly Ile Arg Gln LeuArg Ala Arg Leu Leu Ala Leu Glu Thr 20 25 30 Leu Ile Gln Asn Gln Gln LeuLeu Asn Ser Trp Gly Cys Lys Gly Lys 35 40 45 Ile Val Cys Tyr Thr Ala ValLys Trp Asn Lys Thr Trp Thr Gly Asn 50 55 60 Glu Ser Ile Trp Asp His LeuThr Trp Gln Gln Trp Asp Gln Gln Ile 65 70 75 80 Asp Asn Val Ser Ser ThrIle Tyr Glu Glu Ile Leu Lys Ala Gln Val 85 90 95 Gln Gln Glu Gln Asn GluGln Lys Leu Leu Glu Leu Asp Glu 100 105 110 <210> SEQ ID NO 15 <211>LENGTH: 331 <212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 15aacctgctaa gagcaataca ggcccagcag cacttgctga ggctatctgt atggggtatc 60agacaactcc gagctcgcct gctagcctta gaaaccttaa tacagaatca gcaactccta 120aactcatggg gctgtaaggg aaagatagtc tgttacacag cagtaaaatg gaacaggaca 180tggacaggaa atgaaagtat ttgggaccac ctcacatggc agcaatggga tcagcagata 240gacaatgtaa gctccaccat atatgaggaa atactaaaag cacaagtaca gcaggaacag 300aatgagmaaa arttgctgga gttagatgaa t 331 <210> SEQ ID NO 16 <211> LENGTH:110 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY:misc_feature <222> LOCATION: (1)..(110) <223> OTHER INFORMATION: Xaa =Unknown <400> SEQUENCE: 16 Asn Leu Leu Arg Ala Ile Gln Ala Gln Gln HisLeu Leu Arg Leu Ser 1 5 10 15 Val Trp Gly Ile Arg Gln Leu Arg Ala ArgLeu Leu Ala Leu Glu Thr 20 25 30 Leu Ile Gln Asn Gln Gln Leu Leu Asn SerTrp Gly Cys Lys Gly Lys 35 40 45 Ile Val Cys Tyr Thr Ala Val Lys Trp AsnArg Thr Trp Thr Gly Asn 50 55 60 Glu Ser Ile Trp Asp His Leu Thr Trp GlnGln Trp Asp Gln Gln Ile 65 70 75 80 Asp Asn Val Ser Ser Thr Ile Tyr GluGlu Ile Leu Lys Ala Gln Val 85 90 95 Gln Gln Glu Gln Asn Glu Xaa Xaa LeuLeu Glu Leu Asp Glu 100 105 110 <210> SEQ ID NO 17 <211> LENGTH: 340<212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 17 aacctgctaaaagcaataca ggcccagcag gaattgctga ggctatctgt atggggtatc 60 agacaactccgagctcgcct gctagcctta gaaaccttaa tacaggatca gcagctccta 120 aacctatggggttgtaaggg aaggatagtc tgctacacat cagtaaaatg gaacgataca 180 tggagacatgtcactaatat gagtgaagtt tgggacaaat taacctggca ggaatgggat 240 cggcagatagacaacataag ctatgttata tatgatgaaa tacaaagagc acaagtacag 300 caaggaccaaatgagaagaa gttgctggag ttagatgaat 340 <210> SEQ ID NO 18 <211> LENGTH:113 <212> TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE: 18 Asn Leu LeuLys Ala Ile Gln Ala Gln Gln Glu Leu Leu Arg Leu Ser 1 5 10 15 Val TrpGly Ile Arg Gln Leu Arg Ala Arg Leu Leu Ala Leu Glu Thr 20 25 30 Leu IleGln Asp Gln Gln Leu Leu Asn Leu Trp Gly Cys Lys Gly Arg 35 40 45 Ile ValCys Tyr Thr Ser Val Lys Trp Asn Asp Thr Trp Arg His Val 50 55 60 Thr AsnMet Ser Glu Val Trp Asp Lys Leu Thr Trp Gln Glu Trp Asp 65 70 75 80 ArgGln Ile Asp Asn Ile Ser Tyr Val Ile Tyr Asp Glu Ile Gln Arg 85 90 95 AlaGln Val Gln Gln Gly Pro Asn Glu Lys Lys Leu Leu Glu Leu Asp 100 105 110Glu <210> SEQ ID NO 19 <211> LENGTH: 348 <212> TYPE: DNA <213> ORGANISM:Human <400> SEQUENCE: 19 aacctgctaa gagcaataca ggcccagcag caattgctgaggctatctgt atggggtatc 60 agacaactcc gagctcgcct gctagcctta gaaaccttaatacagaacca gcaactccta 120 aacctatggg gctgtaaggg aaggctagtc tgctacacatcagtaaaatg gaacaagaca 180 tggataaata aaactgacac tgagatagag aatatttgggaaaatctgac atggcaggaa 240 tgggatcagc aaataagcaa cataagctcc accatatatgaggaaataca aaaggcacaa 300 atacaacagg aacataatga gaaaaagttg ctggagctagatgaatgg 348 <210> SEQ ID NO 20 <211> LENGTH: 116 <212> TYPE: PRT <213>ORGANISM: Human <400> SEQUENCE: 20 Asn Leu Leu Arg Ala Ile Gln Ala GlnGln Gln Leu Leu Arg Leu Ser 1 5 10 15 Val Trp Gly Ile Arg Gln Leu ArgAla Arg Leu Leu Ala Leu Glu Thr 20 25 30 Leu Ile Gln Asn Gln Gln Leu LeuAsn Leu Trp Gly Cys Lys Gly Arg 35 40 45 Leu Val Cys Tyr Thr Ser Val LysTrp Asn Lys Thr Trp Ile Asn Lys 50 55 60 Thr Asp Thr Glu Ile Glu Asn IleTrp Glu Asn Leu Thr Trp Gln Glu 65 70 75 80 Trp Asp Gln Gln Ile Ser AsnIle Ser Ser Thr Ile Tyr Glu Glu Ile 85 90 95 Gln Lys Ala Gln Ile Gln GlnGlu His Asn Glu Lys Lys Leu Leu Glu 100 105 110 Leu Asp Glu Trp 115<210> SEQ ID NO 21 <211> LENGTH: 349 <212> TYPE: DNA <213> ORGANISM:Human <400> SEQUENCE: 21 aacctgctaa gagcaataca ggctcagcat caactgctgaagctatctgt atggggtatc 60 agacaactcc gagctcgcct gctagcctta gaaacctttatacagaatca gcaactccta 120 aacctatggg gctgtaaagg aaacctaatc tgctacacatcagtaaaatg gaacgaaaca 180 tggaaaggag ataggacttt tactgacatg gaaaatatttggaacaacct aacatggcag 240 gaatgggatc agcagataag caacataagc tccaccatatatgacgaaat acaaaaggca 300 caagtacagc aggaacaaaa tgagaaaaag ttactagagttaagtgaat 349 <210> SEQ ID NO 22 <211> LENGTH: 116 <212> TYPE: PRT <213>ORGANISM: Human <400> SEQUENCE: 22 Asn Leu Leu Arg Ala Ile Gln Ala GlnHis Gln Leu Leu Lys Leu Ser 1 5 10 15 Val Trp Gly Ile Arg Gln Leu ArgAla Arg Leu Leu Ala Leu Glu Thr 20 25 30 Phe Ile Gln Asn Gln Gln Leu LeuAsn Leu Trp Gly Cys Lys Gly Asn 35 40 45 Leu Ile Cys Tyr Thr Ser Val LysTrp Asn Glu Thr Trp Lys Gly Asp 50 55 60 Arg Thr Phe Thr Asp Met Glu AsnIle Trp Asn Asn Leu Thr Trp Gln 65 70 75 80 Glu Trp Asp Gln Gln Ile SerAsn Ile Ser Ser Thr Ile Tyr Asp Glu 85 90 95 Ile Gln Lys Ala Gln Val GlnGln Glu Gln Asn Glu Lys Lys Leu Leu 100 105 110 Glu Leu Ser Glu 115<210> SEQ ID NO 23 <211> LENGTH: 346 <212> TYPE: DNA <213> ORGANISM:Human <400> SEQUENCE: 23 aacctgctaa gagcaataca ggcccagcag caattgctgaggctatctgt atggggtatc 60 agacaactcc gagctcgcct gctagcctta gaaaccttaatacagagtca gcaactccta 120 aacctgtggg gctgtaaggg aaggctaatc tgctacacctcagtgcattg gaataagaca 180 tggacaaata agacagataa ggatttggag gatatgtgggacaacctaac atggcagcaa 240 tgggatcagc agataagtaa cataagcgcc accatatatgaggaaataca aaaggcacaa 300 gtacaacaag aatacaatga gagaaagttg ttggagttagatgaat 346 <210> SEQ ID NO 24 <211> LENGTH: 115 <212> TYPE: PRT <213>ORGANISM: Human <400> SEQUENCE: 24 Asn Leu Leu Arg Ala Ile Gln Ala GlnGln Gln Leu Leu Arg Leu Ser 1 5 10 15 Val Trp Gly Ile Arg Gln Leu ArgAla Arg Leu Leu Ala Leu Glu Thr 20 25 30 Leu Ile Gln Ser Gln Gln Leu LeuAsn Leu Trp Gly Cys Lys Gly Arg 35 40 45 Leu Ile Cys Tyr Thr Ser Val HisTrp Asn Lys Thr Trp Thr Asn Lys 50 55 60 Thr Asp Lys Asp Leu Glu Asp MetTrp Asp Asn Leu Thr Trp Gln Gln 65 70 75 80 Trp Asp Gln Gln Ile Ser AsnIle Ser Ala Thr Ile Tyr Glu Glu Ile 85 90 95 Gln Lys Ala Gln Val Gln GlnGlu Tyr Asn Glu Arg Lys Leu Leu Glu 100 105 110 Leu Asp Glu 115 <210>SEQ ID NO 25 <211> LENGTH: 340 <212> TYPE: DNA <213> ORGANISM: Human<220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (1)..(340)<223> OTHER INFORMATION: N = any nucleotide <400> SEQUENCE: 25aacctgctaa gagcaataca ggcccagcag caattgctga ggctatctgt atggggtatc 60anacaactcc gagctcgcct gctagcatya gaaaccttaa tacagaatcw gcaactcctg 120aacctatggg gctgtaaggg aakgctagtc tgctacacat cagtamaatg gaacaggaca 180tggacaaaca atactaattt agattcaatt tgggaaaatc taacatggca ggaatgggat 240cagcagataa gcaacataag ctccaccata tatgaagaaa tacaaaaggc acaartacag 300caggaatacr atgagaaaaa gttgctagag ttagatkaat 340 <210> SEQ ID NO 26 <211>LENGTH: 113 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221>NAME/KEY: Misc_feature <222> LOCATION: (1)..(113) <223> OTHERINFORMATION: Xaa = unknown <400> SEQUENCE: 26 Asn Leu Leu Arg Ala IleGln Ala Gln Gln Gln Leu Leu Arg Leu Ser 1 5 10 15 Val Trp Gly Ile XaaGln Leu Arg Ala Arg Leu Leu Ala Xaa Glu Thr 20 25 30 Leu Ile Gln Asn XaaGln Leu Leu Asn Leu Trp Gly Cys Lys Gly Xaa 35 40 45 Leu Val Cys Tyr ThrSer Val Xaa Trp Asn Arg Thr Trp Thr Asn Asn 50 55 60 Thr Asn Leu Asp SerIle Trp Glu Asn Leu Thr Trp Gln Glu Trp Asp 65 70 75 80 Gln Gln Ile SerAsn Ile Ser Ser Thr Ile Tyr Glu Glu Ile Gln Lys 85 90 95 Ala Gln Xaa GlnGln Glu Tyr Xaa Glu Lys Lys Leu Leu Glu Leu Asp 100 105 110 Xaa <210>SEQ ID NO 27 <211> LENGTH: 340 <212> TYPE: DNA <213> ORGANISM: Human<400> SEQUENCE: 27 aacctgctaa gagcaataca ggcccagcag caattgctgaggctatctgt atggggtatc 60 agacaactcc gagctcgcct gttggcctta gaaaccttaatacagaatca gcaactccta 120 aacctatggg gatgtaaggg aaggctaatc tgctacacatcagtacaatg gaacatgaca 180 tggacaaaca attctaatct ggaaacaatt tgggacaacctaacatggca ggaatgggat 240 cagcagataa acagcataag ctctgtcata tatgaggaaatacaaagggc acaagtacag 300 caggaacaaa acgagaaaaa gttgctggag ttagaggaat340 <210> SEQ ID NO 28 <211> LENGTH: 113 <212> TYPE: PRT <213> ORGANISM:Human <400> SEQUENCE: 28 Asn Leu Leu Arg Ala Ile Gln Ala Gln Gln Gln LeuLeu Arg Leu Ser 1 5 10 15 Val Trp Gly Ile Arg Gln Leu Arg Ala Arg LeuLeu Ala Leu Glu Thr 20 25 30 Leu Ile Gln Asn Gln Gln Leu Leu Asn Leu TrpGly Cys Lys Gly Arg 35 40 45 Leu Ile Cys Tyr Thr Ser Val Gln Trp Asn MetThr Trp Thr Asn Asn 50 55 60 Ser Asn Leu Glu Thr Ile Trp Asp Asn Leu ThrTrp Gln Glu Trp Asp 65 70 75 80 Gln Gln Ile Asn Ser Ile Ser Ser Val IleTyr Glu Glu Ile Gln Arg 85 90 95 Ala Gln Val Gln Gln Glu Gln Asn Glu LysLys Leu Leu Glu Leu Glu 100 105 110 Glu <210> SEQ ID NO 29 <211> LENGTH:331 <212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 29 aacctgctgagagcaataca ggcccagcag caattgctga ggctatctgt atggggtatc 60 agacaactccgagctcgcct gctagcctta gaaaccttaa tacagaatca gcaactccta 120 aacctatggggctgtagagg aaggcaagtc tgctacacat cagtaatatg gaatgagaca 180 tggataggaaacgaaaccat ttgggaagaa ctaacatggc aggaatggga tcggcagata 240 agcaacataagctccaccat atatgatgaa atacaaaagg cacaagtaca gcaggaacaa 300 aatgagaaaaaattgctgga gttagatgaa t 331 <210> SEQ ID NO 30 <211> LENGTH: 110 <212>TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE: 30 Asn Leu Leu Arg AlaIle Gln Ala Gln Gln Gln Leu Leu Arg Leu Ser 1 5 10 15 Val Trp Gly IleArg Gln Leu Arg Ala Arg Leu Leu Ala Leu Glu Thr 20 25 30 Leu Ile Gln AsnGln Gln Leu Leu Asn Leu Trp Gly Cys Arg Gly Arg 35 40 45 Gln Val Cys TyrThr Ser Val Ile Trp Asn Glu Thr Trp Ile Gly Asn 50 55 60 Glu Thr Ile TrpGlu Glu Leu Thr Trp Gln Glu Trp Asp Arg Gln Ile 65 70 75 80 Ser Asn IleSer Ser Thr Ile Tyr Asp Glu Ile Gln Lys Ala Gln Val 85 90 95 Gln Gln GluGln Asn Glu Lys Lys Leu Leu Glu Leu Asp Glu 100 105 110 <210> SEQ ID NO31 <211> LENGTH: 334 <212> TYPE: DNA <213> ORGANISM: Human <220>FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (1)..(334) <223>OTHER INFORMATION: N = any nucleotide <400> SEQUENCE: 31 aacctgctgagagcgataca ggcccagcaa cacttgctga ggttatctgt atggggtatt 60 agacaactccgagctcgcct gcaagcctta gaaaccctta tacagaatca gcaacgccta 120 aacctatggggctgtaaggg aaagatgatc tgttacacat cagtaaaatg gaacacatca 180 tggggagactataatgacag tatttggggc aactanacat ggcaacaatg ggaccaagaa 240 ataagcaatgtaagctccat tatatatgac aaaatacaag aagcacagga ccaacaggag 300 aggaatgtaaaagcattgtt ggagctggat gaat 334 <210> SEQ ID NO 32 <211> LENGTH: 111<212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY:Misc_feature <222> LOCATION: (1)..(111) <223> OTHER INFORMATION: Xaa =unknown <400> SEQUENCE: 32 Asn Leu Leu Arg Ala Ile Gln Ala Gln Gln HisLeu Leu Arg Leu Ser 1 5 10 15 Val Trp Gly Ile Arg Gln Leu Arg Ala ArgLeu Gln Ala Leu Glu Thr 20 25 30 Leu Ile Gln Asn Gln Gln Arg Leu Asn LeuTrp Gly Cys Lys Gly Lys 35 40 45 Met Ile Cys Tyr Thr Ser Val Lys Trp AsnThr Ser Trp Gly Asp Tyr 50 55 60 Asn Asp Ser Ile Trp Gly Asn Xaa Thr TrpGln Gln Trp Asp Gln Glu 65 70 75 80 Ile Ser Asn Val Ser Ser Ile Ile TyrAsp Lys Ile Gln Glu Ala Gln 85 90 95 Asp Gln Gln Glu Arg Asn Val Lys AlaLeu Leu Glu Leu Asp Glu 100 105 110 <210> SEQ ID NO 33 <211> LENGTH: 334<212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 33 aacctgctgagagcgataca ggcccagcaa cacttgctga ggttatctgt atggggtatc 60 agacaactccgagctcgcct gcaagcctta gaaaccctta tacagaatca gcaacgccta 120 aacctatggggctgtaaggg aaagatgatc tgttacacat cagtaccatg gaacacatca 180 tggggaaactataatgacag tatttgggat aagtatacat ggcaacaatg ggaccgagaa 240 atagacaatgtaagctacat tatatatgaa aaaatacaag aagcacaaga ccaacaggag 300 aagaatgtaaaagcattgtt ggagctagat gaat 334 <210> SEQ ID NO 34 <211> LENGTH: 111<212> TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE: 34 Asn Leu Leu ArgAla Ile Gln Ala Gln Gln His Leu Leu Arg Leu Ser 1 5 10 15 Val Trp GlyIle Arg Gln Leu Arg Ala Arg Leu Gln Ala Leu Glu Thr 20 25 30 Leu Ile GlnAsn Gln Gln Arg Leu Asn Leu Trp Gly Cys Lys Gly Lys 35 40 45 Met Ile CysTyr Thr Ser Val Pro Trp Asn Thr Ser Trp Gly Asn Tyr 50 55 60 Asn Asp SerIle Trp Asp Lys Tyr Thr Trp Gln Gln Trp Asp Arg Glu 65 70 75 80 Ile AspAsn Val Ser Tyr Ile Ile Tyr Glu Lys Ile Gln Glu Ala Gln 85 90 95 Asp GlnGln Glu Lys Asn Val Lys Ala Leu Leu Glu Leu Asp Glu 100 105 110 <210>SEQ ID NO 35 <211> LENGTH: 331 <212> TYPE: DNA <213> ORGANISM: Human<400> SEQUENCE: 35 aacctgctga gagcaataca ggcccagcaa catctgctgaggttatctgt atggggtatt 60 agacaactcc gagctcgcct gcaagcctta gaaacccttatgcaaaatca gcaactccta 120 aacctatggg gctgtaaagg aaaatcaatc tgctacacatcagtaaaatg gaacaacaca 180 tggggaggaa atctctcaat ttgggacagc ttaacatggcagcaatggga tcaacaggta 240 gccaatgtaa gctctttgat atatgacaaa atacaagaagcacaagaaca acaggaggaa 300 aatgaaaggg ccttgctgga gttagatgaa t 331 <210>SEQ ID NO 36 <211> LENGTH: 110 <212> TYPE: PRT <213> ORGANISM: Human<400> SEQUENCE: 36 Asn Leu Leu Arg Ala Ile Gln Ala Gln Gln His Leu LeuArg Leu Ser 1 5 10 15 Val Trp Gly Ile Arg Gln Leu Arg Ala Arg Leu GlnAla Leu Glu Thr 20 25 30 Leu Met Gln Asn Gln Gln Leu Leu Asn Leu Trp GlyCys Lys Gly Lys 35 40 45 Ser Ile Cys Tyr Thr Ser Val Lys Trp Asn Asn ThrTrp Gly Gly Asn 50 55 60 Leu Ser Ile Trp Asp Ser Leu Thr Trp Gln Gln TrpAsp Gln Gln Val 65 70 75 80 Ala Asn Val Ser Ser Leu Ile Tyr Asp Lys IleGln Glu Ala Gln Glu 85 90 95 Gln Gln Glu Glu Asn Glu Arg Ala Leu Leu GluLeu Asp Glu 100 105 110 <210> SEQ ID NO 37 <211> LENGTH: 331 <212> TYPE:DNA <213> ORGANISM: Human <400> SEQUENCE: 37 aacctgttga gagcgatacaggcccagcaa cacctgctga ggttatctgt atggggtata 60 agacaactcc gagctcgcctgcaagcctta gaaaccttta tacagaacca gcaactccta 120 agcctatggg gatgtaagggaaagctaatc tgttacacat ctgtaaaatg gaacacatca 180 tggggaggaa atgagagtatttggaacaat ctaacatggc agcagtggga tcaacagata 240 gacaacataa gttccatcatatatgatgaa atacaaaagg cacaagagca acaggaacaa 300 aatgagaaaa gcttgctggagttagatgaa t 331 <210> SEQ ID NO 38 <211> LENGTH: 110 <212> TYPE: PRT<213> ORGANISM: Human <400> SEQUENCE: 38 Asn Leu Leu Arg Ala Ile Gln AlaGln Gln His Leu Leu Arg Leu Ser 1 5 10 15 Val Trp Gly Ile Arg Gln LeuArg Ala Arg Leu Gln Ala Leu Glu Thr 20 25 30 Phe Ile Gln Asn Gln Gln LeuLeu Ser Leu Trp Gly Cys Lys Gly Lys 35 40 45 Leu Ile Cys Tyr Thr Ser ValLys Trp Asn Thr Ser Trp Gly Gly Asn 50 55 60 Glu Ser Ile Trp Asn Asn LeuThr Trp Gln Gln Trp Asp Gln Gln Ile 65 70 75 80 Asp Asn Ile Ser Ser IleIle Tyr Asp Glu Ile Gln Lys Ala Gln Glu 85 90 95 Gln Gln Glu Gln Asn GluLys Ser Leu Leu Glu Leu Asp Glu 100 105 110 <210> SEQ ID NO 39 <211>LENGTH: 340 <212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 39aacctgctaa gagcaataca ggcccagcaa gagctgctga ggctatctgt atggggtatc 60agacaactcc gagctcgcct gctagcctta gaaaccttta tacggaatca gcaactccta 120aacctctggg gctgtaaggg aaggctaatt tgctatacat cagtacaatg gaacaaaaca 180tggggtaatt tgamwgataa tgagtcaatt tgggatgaca tracatggca ggagtgggat 240aagcgggtag akaatgtaag ygccaccata tttgaagaaa tacgaagggc acaagaacaa 300caggaacaaa atgagaaggc tttgctagaa ttagatgaat 340 <210> SEQ ID NO 40 <211>LENGTH: 113 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221>NAME/KEY: Misc_feature <222> LOCATION: (1)..(113) <223> OTHERINFORMATION: Xaa = unknown <400> SEQUENCE: 40 Asn Leu Leu Arg Ala IleGln Ala Gln Gln Glu Leu Leu Arg Leu Ser 1 5 10 15 Val Trp Gly Ile ArgGln Leu Arg Ala Arg Leu Leu Ala Leu Glu Thr 20 25 30 Phe Ile Arg Asn GlnGln Leu Leu Asn Leu Trp Gly Cys Lys Gly Arg 35 40 45 Leu Ile Cys Tyr ThrSer Val Gln Trp Asn Lys Thr Trp Gly Asn Leu 50 55 60 Xaa Asp Asn Glu SerIle Trp Asp Asp Xaa Thr Trp Gln Glu Trp Asp 65 70 75 80 Lys Arg Val XaaAsn Val Xaa Ala Thr Ile Phe Glu Glu Ile Arg Arg 85 90 95 Ala Gln Glu GlnGln Glu Gln Asn Glu Lys Ala Leu Leu Glu Leu Asp 100 105 110 Glu <210>SEQ ID NO 41 <211> LENGTH: 315 <212> TYPE: DNA <213> ORGANISM: Human<400> SEQUENCE: 41 taagattatg ggaaaaaata tctcggacag tgcagaaaatatcatagtga ccctaaattc 60 tactgtaaac ataacctgtg agagaccagg gaatcagtcagtacaagaga taaaaatagg 120 tccaatggcc tggtacagca ttggcatagg gacaacacccgcaaactggt caaggatagc 180 ttattgccag tataatatca ctgattggga aaaagccttaaaacaaacag ctgaaaggta 240 cttagaactt gtaaaccata caagaaatga tactgttagcataacattca atagcagcac 300 tggtggagat ctaga 315 <210> SEQ ID NO 42 <211>LENGTH: 103 <212> TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE: 42 IleMet Gly Lys Asn Ile Ser Asp Ser Ala Glu Asn Ile Ile Val Thr 1 5 10 15Leu Asn Ser Thr Val Asn Ile Thr Cys Glu Arg Pro Gly Asn Gln Ser 20 25 30Val Gln Glu Ile Lys Ile Gly Pro Met Ala Trp Tyr Ser Ile Gly Ile 35 40 45Gly Thr Thr Pro Ala Asn Trp Ser Arg Ile Ala Tyr Cys Gln Tyr Asn 50 55 60Ile Thr Asp Trp Glu Lys Ala Leu Lys Gln Thr Ala Glu Arg Tyr Leu 65 70 7580 Glu Leu Val Asn His Thr Arg Asn Asp Thr Val Ser Ile Thr Phe Asn 85 9095 Ser Ser Thr Gly Gly Asp Leu 100 <210> SEQ ID NO 43 <211> LENGTH: 327<212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 43 attataggaaaaaacatttc ggacagtggg aaaaatatca tagtgaccct aaatcctact 60 gtaaacctgacttgtgagag accaggaaat aattcaatac aacagatgaa aataggtcca 120 ctggcctggtacagcatggg cctagagaga aacaaaagct caatctctag attagcttat 180 tgcaggtataataccactac gtgggaacaa gccttacaac aaacagctga aaggtatcta 240 gaacttgtgaacaacacgga caatattaca ataatgttca atcgcagcac tgatggagat 300 tcagaggtaacccatatgca ttttaac 327 <210> SEQ ID NO 44 <211> LENGTH: 109 <212> TYPE:PRT <213> ORGANISM: Human <400> SEQUENCE: 44 Ile Ile Gly Lys Asn Ile SerAsp Ser Gly Lys Asn Ile Ile Val Thr 1 5 10 15 Leu Asn Pro Thr Val AsnLeu Thr Cys Glu Arg Pro Gly Asn Asn Ser 20 25 30 Ile Gln Gln Met Lys IleGly Pro Leu Ala Trp Tyr Ser Met Gly Leu 35 40 45 Glu Arg Asn Lys Ser SerIle Ser Arg Leu Ala Tyr Cys Arg Tyr Asn 50 55 60 Thr Thr Thr Trp Glu GlnAla Leu Gln Gln Thr Ala Glu Arg Tyr Leu 65 70 75 80 Glu Leu Val Asn AsnThr Asp Asn Ile Thr Ile Met Phe Asn Arg Ser 85 90 95 Thr Asp Gly Asp SerGlu Val Thr His Met His Phe Asn 100 105 <210> SEQ ID NO 45 <211> LENGTH:356 <212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 45 ataagaattatgggaaaaaa tatctcgaac agtgcagtaa atatcatagt gaccctgaat 60 tctactgtaaacataacctg tgtgagacca tggaatcaga cagtacaaga gatacaaaca 120 ggtccaatggcctggtatag cattcacttg aggacaccac tcgcaaactt gtcaaggata 180 gcttattgcaagtataatgc cgctgattgg gaaaaagcct taaaacaaac agctgaaagg 240 tacttagaacttgtaaataa tacaagtaat aataatgtta ccataatatt caataacagc 300 actggtggagatccagagac aacccagtta cattttaact gtcatggagt tcttta 356 <210> SEQ ID NO46 <211> LENGTH: 116 <212> TYPE: PRT <213> ORGANISM: Human <400>SEQUENCE: 46 Ile Met Gly Lys Asn Ile Ser Asn Ser Ala Val Asn Ile Ile ValThr 1 5 10 15 Leu Asn Ser Thr Val Asn Ile Thr Cys Val Arg Pro Trp AsnGln Thr 20 25 30 Val Gln Glu Ile Gln Thr Gly Pro Met Ala Trp Tyr Ser IleHis Leu 35 40 45 Arg Thr Pro Leu Ala Asn Leu Ser Arg Ile Ala Tyr Cys LysTyr Asn 50 55 60 Ala Ala Asp Trp Glu Lys Ala Leu Lys Gln Thr Ala Glu ArgTyr Leu 65 70 75 80 Glu Leu Val Asn Asn Thr Ser Asn Asn Asn Val Thr IleIle Phe Asn 85 90 95 Asn Ser Thr Gly Gly Asp Pro Glu Thr Thr Gln Leu HisPhe Asn Cys 100 105 110 His Gly Val Leu 115 <210> SEQ ID NO 47 <211>LENGTH: 317 <212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 47ataagactga tggcaaaaaa tatttcggct actggccaaa atatcatagt gaccctaaat 60actactataa acatgacctg ccagagacca ggaaatctaa caatacagga aataaagata 120ggtccaatgt cctggtacag catgggcata gggcaggaag accactctaa gtcaagaaac 180gcttattgtg agtataatat cactgattgg gtacaggcct taaaacagac agctgaaagg 240tatttagaat tagtaaacaa tacaaatact aatataaaca tgacattcga gaacagtact 300ggaggagatc cagaggt 317 <210> SEQ ID NO 48 <211> LENGTH: 103 <212> TYPE:PRT <213> ORGANISM: Human <400> SEQUENCE: 48 Leu Met Ala Lys Asn Ile SerAla Thr Gly Gln Asn Ile Ile Val Thr 1 5 10 15 Leu Asn Thr Thr Ile AsnMet Thr Cys Gln Arg Pro Gly Asn Leu Thr 20 25 30 Ile Gln Glu Ile Lys IleGly Pro Met Ser Trp Tyr Ser Met Gly Ile 35 40 45 Gly Gln Glu Asp His SerLys Ser Arg Asn Ala Tyr Cys Glu Tyr Asn 50 55 60 Ile Thr Asp Trp Val GlnAla Leu Lys Gln Thr Ala Glu Arg Tyr Leu 65 70 75 80 Glu Leu Val Asn AsnThr Asn Thr Asn Ile Asn Met Thr Phe Glu Asn 85 90 95 Ser Thr Gly Gly AspPro Glu 100 <210> SEQ ID NO 49 <211> LENGTH: 319 <212> TYPE: DNA <213>ORGANISM: Human <400> SEQUENCE: 49 taagaataat gggaaaaaat atttcagacaatgggaaaaa tatcatagta accctaaatt 60 ctactctaaa aatgacctgt gagagaccagggaatcatac agtacaacag atgaagatag 120 gtccaatgtc ctggtatagc atgggcttagagaaaaacaa taccagctca agaagagctt 180 tttgcaagta taatgccact aattgggaaaaaaccttaaa acaaatggct gaaaggtatt 240 tagaactcgt aaacaataca agtaataacacagtgacaat gatattcaat acaagcagtg 300 atggagatcc agaggtacc 319 <210> SEQID NO 50 <211> LENGTH: 104 <212> TYPE: PRT <213> ORGANISM: Human <400>SEQUENCE: 50 Ile Met Gly Lys Asn Ile Ser Asp Asn Gly Lys Asn Ile Ile ValThr 1 5 10 15 Leu Asn Ser Thr Leu Lys Met Thr Cys Glu Arg Pro Gly AsnHis Thr 20 25 30 Val Gln Gln Met Lys Ile Gly Pro Met Ser Trp Tyr Ser MetGly Leu 35 40 45 Glu Lys Asn Asn Thr Ser Ser Arg Arg Ala Phe Cys Lys TyrAsn Ala 50 55 60 Thr Asn Trp Glu Lys Thr Leu Lys Gln Met Ala Glu Arg TyrLeu Glu 65 70 75 80 Leu Val Asn Asn Thr Ser Asn Asn Thr Val Thr Met IlePhe Asn Thr 85 90 95 Ser Ser Asp Gly Asp Pro Glu Val 100 <210> SEQ ID NO51 <211> LENGTH: 331 <212> TYPE: DNA <213> ORGANISM: Human <400>SEQUENCE: 51 aagaaggatg ggggaaaaca atccttcaga tcggaagaag atcctagtgaccctaaattc 60 ccctataaac ataacctgcg agagaccata ctatcagtca gtacaagagttaaggatagg 120 tccaatggct tggtacagca tgacattaga acgagacagg gcaggcagtgacataagggc 180 agcttattgc aagtataatg cctctgactg gagaaataca ttaaaaggagtagctgagag 240 atatttagaa cttagaaatg aggaaggccc ggtgaacgtg accttcaatggaagtgcggg 300 tggagatcca gagatacgct ttctgcattt t 331 <210> SEQ ID NO 52<211> LENGTH: 109 <212> TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE:52 Arg Met Gly Glu Asn Asn Pro Ser Asp Arg Lys Lys Ile Leu Val Thr 1 510 15 Leu Asn Ser Pro Ile Asn Ile Thr Cys Glu Arg Pro Tyr Tyr Gln Ser 2025 30 Val Gln Glu Leu Arg Ile Gly Pro Met Ala Trp Tyr Ser Met Thr Leu 3540 45 Glu Arg Asp Arg Ala Gly Ser Asp Ile Arg Ala Ala Tyr Cys Lys Tyr 5055 60 Asn Ala Ser Asp Trp Arg Asn Thr Leu Lys Gly Val Ala Glu Arg Tyr 6570 75 80 Leu Glu Leu Arg Asn Glu Glu Gly Pro Val Asn Val Thr Phe Asn Gly85 90 95 Ser Ala Gly Gly Asp Pro Glu Ile Arg Phe Leu His Phe 100 105<210> SEQ ID NO 53 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Human<220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(6) <400>SEQUENCE: 53 Val Gln Gln Met Lys Ile 1 5 <210> SEQ ID NO 54 <211>LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221>NAME/KEY: PEPTIDE <222> LOCATION: (1)..(11) <400> SEQUENCE: 54 Lys IleGly Pro Met Ser Trp Tyr Ser Met Gly 1 5 10 <210> SEQ ID NO 55 <211>LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221>NAME/KEY: PEPTIDE <222> LOCATION: (1)..(6) <400> SEQUENCE: 55 Met GlyLeu Glu Lys Asn 1 5 <210> SEQ ID NO 56 <211> LENGTH: 6 <212> TYPE: PRT<213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222>LOCATION: (1)..(6) <400> SEQUENCE: 56 Ile Gln Gln Met Lys Ile 1 5 <210>SEQ ID NO 57 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Human<220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(11) <400>SEQUENCE: 57 Lys Ile Gly Pro Leu Ala Trp Tyr Ser Met Gly 1 5 10 <210>SEQ ID NO 58 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Human <220>FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(6) <400>SEQUENCE: 58 Met Gly Leu Glu Arg Asn 1 5 <210> SEQ ID NO 59 <211>LENGTH: 8 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221>NAME/KEY: PEPTIDE <222> LOCATION: (1)..(8) <400> SEQUENCE: 59 Gln SerVal Gln Glu Ile Lys Ile 1 5 <210> SEQ ID NO 60 <211> LENGTH: 11 <212>TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE<222> LOCATION: (1)..(11) <400> SEQUENCE: 60 Lys Ile Gly Pro Met Ala TrpTyr Ser Ile Gly 1 5 10 <210> SEQ ID NO 61 <211> LENGTH: 6 <212> TYPE:PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222>LOCATION: (1)..(6) <400> SEQUENCE: 61 Ile Gly Ile Gly Thr Thr 1 5 <210>SEQ ID NO 62 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Human <220>FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(6) <400>SEQUENCE: 62 Val Gln Glu Ile Gln Thr 1 5 <210> SEQ ID NO 63 <211>LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221>NAME/KEY: PEPTIDE <222> LOCATION: (1)..(11) <400> SEQUENCE: 63 Gln ThrGly Pro Met Ala Trp Tyr Ser Ile His 1 5 10 <210> SEQ ID NO 64 <211>LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221>NAME/KEY: PEPTIDE <222> LOCATION: (1)..(6) <400> SEQUENCE: 64 Ile HisLeu Arg Thr Pro 1 5 <210> SEQ ID NO 65 <211> LENGTH: 6 <212> TYPE: PRT<213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222>LOCATION: (1)..(6) <400> SEQUENCE: 65 Ile Gln Glu Ile Lys Ile 1 5 <210>SEQ ID NO 66 <211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM: Human<220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(11) <400>SEQUENCE: 66 Lys Ile Gly Pro Met Ser Trp Tyr Ser Met Gly 1 5 10 <210>SEQ ID NO 67 <211> LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Human <220>FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(6) <400>SEQUENCE: 67 Met Gly Ile Gly Gln Glu 1 5 <210> SEQ ID NO 68 <211>LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221>NAME/KEY: PEPTIDE <222> LOCATION: (1)..(7) <400> SEQUENCE: 68 Ser ValGln Glu Leu Arg Ile 1 5 <210> SEQ ID NO 69 <211> LENGTH: 11 <212> TYPE:PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222>LOCATION: (1)..(11) <400> SEQUENCE: 69 Arg Ile Gly Pro Met Ala Trp TyrSer Met Thr 1 5 10 <210> SEQ ID NO 70 <211> LENGTH: 6 <212> TYPE: PRT<213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222>LOCATION: (1)..(6) <400> SEQUENCE: 70 Met Thr Leu Glu Arg Asp 1 5 <210>SEQ ID NO 71 <211> LENGTH: 17 <212> TYPE: PRT <213> ORGANISM: Human<220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(17) <400>SEQUENCE: 71 Arg Asn Gln Gln Leu Leu Asn Leu Trp Gly Cys Lys Gly Arg LeuIle 1 5 10 15 Cys <210> SEQ ID NO 72 <211> LENGTH: 15 <212> TYPE: PRT<213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222>LOCATION: (1)..(15) <400> SEQUENCE: 72 Cys Lys Gly Arg Leu Ile Cys TyrThr Ser Val Gln Trp Asn Met 1 5 10 15 <210> SEQ ID NO 73 <211> LENGTH:10 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY:PEPTIDE <222> LOCATION: (1)..(10) <400> SEQUENCE: 73 Leu Trp Gly Cys LysGly Arg Ile Val Cys 1 5 10 <210> SEQ ID NO 74 <211> LENGTH: 11 <212>TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE<222> LOCATION: (1)..(11) <400> SEQUENCE: 74 Ser Leu Trp Gly Cys Lys GlyLys Leu Ile Cys 1 5 10 <210> SEQ ID NO 75 <211> LENGTH: 7 <212> TYPE:PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222>LOCATION: (1)..(7) <400> SEQUENCE: 75 Cys Lys Gly Lys Ser Ile Cys 1 5<210> SEQ ID NO 76 <211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Human<220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(7) <400>SEQUENCE: 76 Cys Lys Gly Lys Ile Val Cys 1 5 <210> SEQ ID NO 77 <211>LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221>NAME/KEY: PEPTIDE <222> LOCATION: (1)..(7) <400> SEQUENCE: 77 Cys ArgGly Arg Gln Val Cys 1 5 <210> SEQ ID NO 78 <400> SEQUENCE: 78 000 <210>SEQ ID NO 79 <211> LENGTH: 12 <212> TYPE: PRT <213> ORGANISM: Human<220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(12) <400>SEQUENCE: 79 Cys Lys Gly Arg Leu Ile Cys Tyr Thr Ser Val His 1 5 10<210> SEQ ID NO 80 <211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Human<220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(7) <400>SEQUENCE: 80 Cys Lys Gly Asn Leu Ile Cys 1 5 <210> SEQ ID NO 81 <211>LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221>NAME/KEY: PEPTIDE <222> LOCATION: (1)..(7) <400> SEQUENCE: 81 Cys LysGly Lys Met Ile Cys 1 5 <210> SEQ ID NO 82 <211> LENGTH: 7 <212> TYPE:PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222>LOCATION: (1)..(7) <400> SEQUENCE: 82 Cys Lys Gly Arg Val Val Cys 1 5<210> SEQ ID NO 83 <211> LENGTH: 37 <212> TYPE: PRT <213> ORGANISM:Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(37)<400> SEQUENCE: 83 Cys Glu Arg Pro Gly Asn Asn Ser Ile Gln Gln Met LysIle Gly Pro 1 5 10 15 Leu Ala Trp Tyr Ser Met Gly Leu Glu Arg Asn LysSer Ser Ile Ser 20 25 30 Arg Leu Ala Tyr Cys 35 <210> SEQ ID NO 84 <211>LENGTH: 37 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221>NAME/KEY: PEPTIDE <222> LOCATION: (1)..(37) <400> SEQUENCE: 84 Cys GluArg Pro Gly Asn Asn Ser Ile Gln Gln Met Lys Ile Gly Pro 1 5 10 15 MetAla Trp Tyr Ser Met Gly Leu Glu Arg Asn Lys Ser Ser Ile Ser 20 25 30 ArgLeu Ala Tyr Cys 35 <210> SEQ ID NO 85 <211> LENGTH: 37 <212> TYPE: PRT<213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222>LOCATION: (1)..(37) <400> SEQUENCE: 85 Cys Glu Arg Pro Gly Asn Gln SerVal Gln Glu Ile Lys Ile Gly Pro 1 5 10 15 Met Ala Trp Tyr Ser Ile GlyIle Gly Thr Thr Pro Ala Asn Trp Ser 20 25 30 Arg Ile Ala Tyr Cys 35<210> SEQ ID NO 86 <211> LENGTH: 38 <212> TYPE: PRT <213> ORGANISM:Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(38)<400> SEQUENCE: 86 Cys Glu Arg Pro Gly Asn Gln Ser Val Gln Glu Ile LysIle Gly Pro 1 5 10 15 Met Ala Trp Tyr Ser Ile Gly Ile Gly Thr Thr ProThr Tyr Asn Trp 20 25 30 Ser Arg Ile Ala Tyr Cys 35 <210> SEQ ID NO 87<211> LENGTH: 37 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE:<221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(37) <400> SEQUENCE: 87 CysVal Arg Pro Trp Asn Gln Thr Val Gln Glu Ile Gln Thr Gly Pro 1 5 10 15Met Ala Trp Tyr Ser Ile His Leu Arg Thr Pro Leu Ala Asn Leu Ser 20 25 30Arg Ile Ala Tyr Cys 35 <210> SEQ ID NO 88 <211> LENGTH: 37 <212> TYPE:PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222>LOCATION: (1)..(37) <400> SEQUENCE: 88 Cys Gln Arg Pro Gly Asn Leu ThrIle Gln Glu Ile Lys Ile Gly Pro 1 5 10 15 Met Ser Trp Tyr Ser Met GlyIle Gly Gln Glu Asp His Ser Lys Ser 20 25 30 Arg Asn Ala Tyr Cys 35<210> SEQ ID NO 89 <211> LENGTH: 38 <212> TYPE: PRT <213> ORGANISM:Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(38)<400> SEQUENCE: 89 Cys Glu Arg Pro Tyr Tyr Gln Ser Val Gln Glu Leu ArgIle Gly Pro 1 5 10 15 Met Ala Trp Tyr Ser Met Thr Leu Glu Arg Asp ArgAla Gly Ser Asp 20 25 30 Ile Arg Ala Ala Tyr Cys 35 <210> SEQ ID NO 90<211> LENGTH: 36 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE:<221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(36) <400> SEQUENCE: 90 CysGlu Arg Pro Gly Asn His Thr Val Gln Gln Met Lys Ile Gly Pro 1 5 10 15Met Ser Trp Tyr Ser Met Gly Leu Glu Lys Asn Asn Thr Ser Ser Arg 20 25 30Arg Ala Phe Cys 35 <210> SEQ ID NO 91 <211> LENGTH: 23 <212> TYPE: PRT<213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222>LOCATION: (1)..(23) <400> SEQUENCE: 91 Asp Gln Gln Leu Leu Asn Leu TrpGly Cys Lys Gly Arg Ile Val Cys 1 5 10 15 Tyr Thr Ser Val Lys Trp Asn 20<210> SEQ ID NO 92 <211> LENGTH: 24 <212> TYPE: PRT <213> ORGANISM:Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(24)<400> SEQUENCE: 92 Asn Gln Gln Leu Leu Asn Leu Trp Gly Cys Lys Gly ArgLeu Val Cys 1 5 10 15 Tyr Thr Ser Val Lys Trp Asn Lys 20 <210> SEQ ID NO93 <211> LENGTH: 23 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE:<221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(23) <400> SEQUENCE: 93 AsnGln Gln Arg Leu Asn Leu Trp Gly Cys Lys Gly Lys Met Ile Cys 1 5 10 15Tyr Thr Ser Val Pro Trp Asn 20 <210> SEQ ID NO 94 <211> LENGTH: 23 <212>TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE<222> LOCATION: (1)..(23) <400> SEQUENCE: 94 Asn Gln Gln Leu Leu Asn LeuTrp Gly Cys Lys Gly Lys Ser Ile Cys 1 5 10 15 Tyr Thr Ser Val Lys TrpAsn 20 <210> SEQ ID NO 95 <211> LENGTH: 23 <212> TYPE: PRT <213>ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION:(1)..(23) <400> SEQUENCE: 95 Asn Gln Gln Leu Leu Asn Leu Trp Gly Cys LysGly Arg Leu Ile Cys 1 5 10 15 Tyr Thr Ser Val Gln Trp Asn 20 <210> SEQID NO 96 <211> LENGTH: 23 <212> TYPE: PRT <213> ORGANISM: Human <220>FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(23) <400>SEQUENCE: 96 Asn Gln Gln Arg Leu Asn Leu Trp Gly Cys Lys Gly Lys Met IleCys 1 5 10 15 Tyr Thr Ser Val Lys Trp Asn 20 <210> SEQ ID NO 97 <211>LENGTH: 23 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221>NAME/KEY: PEPTIDE <222> LOCATION: (1)..(23) <400> SEQUENCE: 97 Asn GlnGln Leu Leu Asn Leu Trp Gly Cys Lys Gly Asn Leu Ile Cys 1 5 10 15 TyrThr Ser Val Lys Trp Asn 20 <210> SEQ ID NO 98 <211> LENGTH: 23 <212>TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE<222> LOCATION: (1)..(23) <400> SEQUENCE: 98 Asn Gln Gln Leu Leu Asn LeuTrp Gly Cys Arg Gly Arg Gln Val Cys 1 5 10 15 Tyr Thr Ser Val Ile TrpAsn 20 <210> SEQ ID NO 99 <211> LENGTH: 23 <212> TYPE: PRT <213>ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION:(1)..(23) <400> SEQUENCE: 99 Ser Gln Gln Leu Leu Asn Leu Trp Gly Cys LysGly Arg Leu Ile Cys 1 5 10 15 Tyr Thr Ser Val His Trp Asn 20 <210> SEQID NO 100 <211> LENGTH: 23 <212> TYPE: PRT <213> ORGANISM: Human <220>FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(23) <400>SEQUENCE: 100 Asn Gln Gln Leu Leu Asn Leu Trp Gly Cys Lys Gly Arg IleVal Cys 1 5 10 15 Tyr Thr Ser Val Lys Trp Asn 20 <210> SEQ ID NO 101<211> LENGTH: 23 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE:<221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(23) <400> SEQUENCE: 101Asn Gln Gln Leu Leu Asn Ser Trp Gly Cys Lys Gly Lys Ile Val Cys 1 5 1015 Tyr Thr Ala Val Lys Trp Asn 20 <210> SEQ ID NO 102 <211> LENGTH: 23<212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY:PEPTIDE <222> LOCATION: (1)..(23) <400> SEQUENCE: 102 Asn Gln Gln LeuLeu Ser Leu Trp Gly Cys Lys Gly Lys Leu Ile Cys 1 5 10 15 Tyr Thr SerVal Lys Trp Asn 20 <210> SEQ ID NO 103 <211> LENGTH: 1339 <212> TYPE:DNA <213> ORGANISM: Human <400> SEQUENCE: 103 attaaagtag taccaagaagaaaggcaaaa ataatcagac attatggaaa acagatggca 60 ggtgctgata gtatggcaagtggacagaca gaaagtgaaa gcgtggaaca gcctggtgaa 120 ataccataag tacaggtctaggaaggccaa ggactggtgt tacagacacc attttgaatc 180 tagaaatcca agagtcagttcaagtgtaca tattccagta gggatggctt gggtaatagt 240 gaccacatat tggggattgatgccagggga gagagaggaa cagttgggac atggggttag 300 tatagaatgg cagtacaaaaagtatacaac acagattgac cctgaaacag cagacaggat 360 gatacatctg tattattttacctgttttac agattcagca gtcaggaaag ccatcttagg 420 gcagagaata ctgaccaagtgtgaataccc tgcaggacat agtcaggtag ggacattgca 480 actactagct ctaagagtagtagtaaaagc aaaaagaaat aagcctcccc tacccagtgt 540 ccagaaatta acagaagatagatggagcga gcacctgagg atcaggggcc agctagagag 600 cctttcaatg aatgggcactagagatccta gaagagctaa aagcagaggc agtaagacat 660 ttccctaggc agtggctacaggccttggga cagtacattt atgagactta tggggacact 720 tgggtaggag ttatggcaattacaagaatc ttacaacaaa tactatttgc ccattttaga 780 attggatgtc aacatagtagaataggaatt aacccaacta atacaagagg aagaggaaga 840 agaaatggat ccagtagatcctgagatgcc cccttggcat caccctggga gtcagcccca 900 gatcccttgt aacaattgctattgcaaaag atgctgctat cattgccttg tttgtttcac 960 aagaaagggt ttggggatctcctatggcag gaagaagcgg cgacaacgaa gagctgctgc 1020 gagccatccg gataataaagatcttgtacc agagcagtaa gtaacgctaa tgcatcatag 1080 ggacctgcta gtattaataattattagtgc tttgctgctt ataaatgtaa ttatatggat 1140 gtttattctt agacaatatttagaacagaa gaaacaggac agaagggaaa gagacatact 1200 tgaaaggtta agaagaatagcagaaattaa agatgatagt gactatgaaa gcaatgaaga 1260 ggaggaacag gaagttagagatcttataca tagtcatggc tttgataatc ccatgtttga 1320 gctctgatca gaagtatgc1339 <210> SEQ ID NO 104 <211> LENGTH: 1282 <212> TYPE: DNA <213>ORGANISM: Human <400> SEQUENCE: 104 attaaagtag taccaagaag aaaggcaaaaataatcagag attatggaaa acaaatggca 60 ggtactgata gtatggcaag tagacagacagaaagtgaaa acgtggaaca gcttggtgaa 120 ataccataag tacaggtcta ggaaggccaaggactggtac tacagacatc attatgaatc 180 tagaaatcca agaatcagtt caggtgtatatattccagta gggccggctt gtatagtagt 240 gaacacatat tggggattga tgccaggagaaagagatgaa catctgggac atggggttag 300 tatagaatgg cagtacaaga agtatacaacacagattgac cctgaaacag cagacaggat 360 gatacatcta tactatttta cctgttttacagaatcagca atcaggaaag ccatcctagg 420 gcagagagta ctgaccaagt gtgaataccctgcaggacat agccaggtag ggacactaca 480 actactagct ctaagagttg tagtaaaagagagaaaacat aggcctcccc tacccagtgt 540 ccagaaatta acagaagata gatggaacaagcacctgagg atcagggacc agctagagag 600 ccattcaatg aatggacact agagctcctagaagagctaa aagcagaagc agtaagacat 660 tttcctaggc cttggctaca ggccttgggacaatacattt atgatactta tggggacact 720 tgggtaggag ttatggcaat tataagactcttacaattaa tgatatttgc ccattttaga 780 agaaatggat ccagtagacc ctgagatgcccccttggcat caccctggaa gtcagcccca 840 gaatccttgt aataaatgct attgcaaaaaatgctgctat cattgctatg tttgtttcac 900 aagcaagggt ttgggaatct cctatggcaggaagaagcga cgacgaccag cagctgctgc 960 aagccgcccg gataataaag atcttgtaccagagcagtaa gtaacgctaa tgcagcaaaa 1020 ggacctgcta ttattagtaa ttattagtgctttgctgctt ataaatataa ttctatggat 1080 gtttaatctt agaaaatatt tagaacaaaagaaacaagac agaagggaaa gagaaatact 1140 tgaaaggata agaagaataa gagaaattagagatgatagt gactatgaaa gcaatgaaga 1200 ggaagaacaa gaagttaggg gtcatcttgtgcatatgttt ggctttgcta atcccgtgtt 1260 tgagatctaa tgacctatat gc 1282<210> SEQ ID NO 105 <211> LENGTH: 1339 <212> TYPE: DNA <213> ORGANISM:Human <400> SEQUENCE: 105 atcaaggtag taccaagaag aaaagcaaaa atactcagggattatggaaa acagatggca 60 ggtgctgata gtatggcaag tggacagaca gaaagtgaaagcatggaata gcctggtaaa 120 ataccataag tacaggtcta gaaagaccca gaactgggattatagacatc attatgaaat 180 cagaaatcca agaatcagct caggtgtata tattccagtaggtgaagcta agatagtagt 240 gactacatat tggggattaa tgccagggga aagagatgagcatttgggac atggagtcag 300 tatagaatgg caatacaaaa attatagtac acagattgaccctgaaacag cagataaaat 360 aatacatctg cattatttca cctgttttac agagtcagcaatcaggagag ccattttagg 420 gcagagagtg ctgaccaggt gtgaataccc tgcaggacatagtcaggtag ggacactgca 480 actcctagca ttaagagcag tagtaaaaga caaaagaagtaaacctcccc tacccagtgt 540 ccagaagtta acaggagaca gatggaacag gcacctgagaatcagggacc agcaagagag 600 ccattcaatg aatgggcatt agagaccctg gaagaaataaaagcagaagc agtaagacac 660 tttcctaggc cttggctaca aagcttagga caatacatctatgagactta tggagacacc 720 tgggaaggag ttatggcaat cataagaatc ttacaacagttgatatttgc ccattttaga 780 attggatgcc aacatagtag aataggaatt accccatctaacgcaagagg aagaggaaga 840 agaaatggat ccagtagatc ctgaggtgcc cccctggcatcaccctggaa gtcagccccc 900 aaccccttgc aacgcttgct attgcaaaag atgctgttatcattgctatc tttgtttcac 960 aaagaagggt ttgggaatct cccatggcag gaagaagcgacgacgaccag cagctgctgc 1020 aagctcttcg aataataaag atcttgtacc agagcagtaagtaaagctaa tgcatcataa 1080 ggacttgcta atcttaatag ttgctagtat tttgctttttacaaatatag tgatatggac 1140 atttattctt aagaaatatt tagagcagaa ggaacaagatagaagggaaa gagaactact 1200 gaaaagaata aaaagaataa gagaagtcag ggatgatagtgattatgaaa gcaatggaga 1260 tggaggacaa gaagttatac atcttgtgca tactcatggttttgttaacc ccatgtttga 1320 gctctgacaa gctatatcg 1339 <210> SEQ ID NO 106<211> LENGTH: 3600 <212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE:106 ttcacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaataatag 60acataatagc atcagatata caaactaaag aactacaaaa acaaattaca aaaattcaaa 120attttcgggt ttattacagg gacagcagag atccaatttg gaaaggacca gcaaaactac 180tctggaaagg tgaaggggca gtagtaatac aggacaatag tgatataaag gtagtaccaa 240gaagaaaagc aaaaatcatt aaggattatg gaaaacagat ggcaggtgat gattgtgtgg 300caagtagaca gaatgaggat tagaacatgg aacagtctag taaagcatca tatgtatatt 360tctaagaaag ctacagattg ggtttataaa catcactatg atagtagaca tccaaaagta 420agctcagaag tacacattcc actaggggat gctaaattgg taataagaac atattggggt 480ctacatacag gagaaagaga ctggcatttg ggtcatgggg tctccataga atggaaacag 540agaagatata gcacacaaat agatcctgac ctagcagacc aactgattca cctgcattat 600tttaactgtt tttcagaatc tgccataaga aaagccatac taggacaagt agttagacct 660aggtgtgatt atccagcagg acatagtaag gtaggatctc tacaatattt ggcactgaaa 720gcattagtaa caccaacaag gacaaagcca cctttgccta gtgttaagaa attaacagaa 780gacagatgga acaagcccca gaagaccagg gggcacagag ggagcggtcc aatgtatgga 840cattagatct attagaggag cttaaacatg aagctgttag acattttcct aggccttggc 900tccagggatt aggacaatat atctatgaaa catatgggga tacctgggaa ggagttgaag 960ctataataag aattttgcaa caactactgt ttgcccattt tagaattgga tgccaacata 1020gtaggatagg aattaaccca tctaacccaa gaggaaaagg aagaagaaat ggatccagta 1080gatcctgaga tacccccttg gcatcaccct ggaagtcagc cccagacccc ttgtaataac 1140tgctcttgca aaaaatgctg ctaccattgc tatgtgtgtt tcacaagaaa gggtttggaa 1200atctcctatg gcaggaagaa gcgacgaaga tcagccgctg aaacgcgtca tccagataat 1260caagatattg taccagagca gtaagtaacg ctaatgcagc ttagggacca gctaacatta 1320ataattatta gtgctttgtt gcttgtaaat gtagttctat ggacatttat tcttagacaa 1380tatttaaagc aaaagaaaca agatagaagg ggaagagaaa tacttgaaag gttaagaaga 1440ataagacaaa ttgaagatga cagtgactat gaaagcgatg gaaaagagga acaggaagtt 1500agggatcttg tgcatagtta tggctttgat aaccccatgt ttgagccatg accaacgcta 1560tgcaacagtg tatgctgggg tacctgtatg ggaagaggca aacccagtat tattttgtgc 1620ttcagatgct aacctaacaa gcactgagaa gcataatatc tgggcatcac aagcctgtgt 1680tcccacagac cccactccac atgaatatcc tttacacaat gtgacagata actttaatat 1740atggaaaaat tacatggtag aacaaatgca ggatgacatt attagcttat gggaacagag 1800tttaaaacct tgtgttcaaa tgactttcct gtgtgtacaa atgaattgta caagtgtaag 1860taatagtagt gtaagtaata gtagtgtaag taatagtagt gtaagtaata gtagtgtaag 1920tgatagtact atacccaaga agaaaaataa cagcagctca gaggaccttc tgaaacagtg 1980tgattttaat gcaaccacag ttctcaaaga caaaaaggag aaaaaacaga ctctatttta 2040tgtatcagat ttgatgaaac tgacaaatgt cacaaatgac acaatgtata cattaattaa 2100ttgtaactcc acaaccatta agcaagcctg tccaaaggta acttttgagc caattccaat 2160acactattgt gctccagcgg ggtatgccat ctttaagtgt aacaacacag agtttaatgg 2220aacgggccca tgcaacaaca ttacagtagt tacttgtaca catggtatca ggccaacagt 2280gagtacgcaa ctaatattaa acgggacact ctctgaagga aaaataagaa ttatgggaag 2340aaatatcacg gacagtggaa aaaatattat agttacccta aattatacta taaacataac 2400ttgtgagaga acatggaatc agtcagtaca agagatacct ataggtccaa tggcctggta 2460cagcatgagc gtagagaaag acaaaaacac aactggctcg aggtcagcag attgccagta 2520taacacctct gaatggacaa gagccttaga acaaacagct gaaaggtatt tagaactgat 2580gaacaataca ggtaatactg ataatactac agtgatattc aatcatagca ctggtggaga 2640tccagaggta tccttcctac attttaattg tcatggagag ttcttctatt gtaacacatc 2700tgggatgttt aattatacct tttcatgtaa aggaactaac tgtacccaag ttggttccca 2760aaatgaatat aataatcata caaccaagat accttgcagg ataaaacagg tggtaaggtc 2820atggataagg ggagggtcgg gactctatgc acctcccagg caaggtcccc taaaatgtag 2880ctcaaacata actggaatga ttctacaatt ggataagcca tggaacagaa gtgggcacaa 2940caatgacacc acatttagac caataggagg agaaatgaaa gatatatgga gaactgaatt 3000gttcaaatac aaagtagtaa aggtaaaacc ttttagtgtg gcacctacaa aaattgcaag 3060gccagtcata ggcacgggca ctcaaagaga aaagagagca gtaggattgg gaatgctatt 3120cttaggggtt ctaagtgcag caggtagcac tatgggcgca gcggcaacaa cgctggcggt 3180acagacccac actttgatga agggtatagt gcaacagcag gacaacctgc taagagcaat 3240acaggcccag cagcaattgc tgaggctatc tgtatggggt atcagacaac tccgagctcg 3300cctgctagca ttagaaacct taatacagaa tcagcaactc ctgaacctat ggggctgtaa 3360gggaaggcta gtctgctaca catcagtaca atggaacagg acatggacaa acaatactaa 3420tttagattca atttgggaaa atctaacatg gcaggaatgg gatcagcaga taagcaacat 3480aagctccacc atatatgagg aaatacaaaa ggcacaaata cagcaggaat acaatgagaa 3540aaagttgcta gagttagatg aatgggcttc tatttggaat tggcttgaca taactaaatg 3600<210> SEQ ID NO 107 <211> LENGTH: 192 <212> TYPE: PRT <213> ORGANISM:Human <400> SEQUENCE: 107 Met Glu Asn Arg Trp Gln Val Leu Ile Val TrpGln Val Asp Arg Gln 1 5 10 15 Lys Val Lys Ala Trp Asn Ser Leu Val LysTyr His Lys Tyr Arg Ser 20 25 30 Arg Lys Ala Lys Asp Trp Cys Tyr Arg HisHis Phe Glu Ser Arg Asn 35 40 45 Pro Arg Val Ser Ser Ser Val His Ile ProVal Gly Met Ala Trp Val 50 55 60 Ile Val Thr Thr Tyr Trp Gly Leu Met ProGly Glu Arg Glu Glu Gln 65 70 75 80 Leu Gly His Gly Val Ser Ile Glu TrpGln Tyr Lys Lys Tyr Thr Thr 85 90 95 Gln Ile Asp Pro Glu Thr Ala Asp ArgMet Ile His Leu Tyr Tyr Phe 100 105 110 Thr Cys Phe Thr Asp Ser Ala ValArg Lys Ala Ile Leu Gly Gln Arg 115 120 125 Ile Leu Thr Lys Cys Glu TyrPro Ala Gly His Ser Gln Val Gly Thr 130 135 140 Leu Gln Leu Leu Ala LeuArg Val Val Val Lys Ala Lys Arg Asn Lys 145 150 155 160 Pro Pro Leu ProSer Val Gln Lys Leu Thr Glu Asp Arg Trp Ser Glu 165 170 175 His Leu ArgIle Arg Gly Gln Leu Glu Ser Leu Ser Met Asn Gly His 180 185 190 <210>SEQ ID NO 108 <211> LENGTH: 192 <212> TYPE: PRT <213> ORGANISM: Human<400> SEQUENCE: 108 Met Glu Asn Lys Trp Gln Val Leu Ile Val Trp Gln ValAsp Arg Gln 1 5 10 15 Lys Val Lys Thr Trp Asn Ser Leu Val Lys Tyr HisLys Tyr Arg Ser 20 25 30 Arg Lys Ala Lys Asp Trp Tyr Tyr Arg His His TyrGlu Ser Arg Asn 35 40 45 Pro Arg Ile Ser Ser Gly Val Tyr Ile Pro Val GlyPro Ala Cys Ile 50 55 60 Val Val Asn Thr Tyr Trp Gly Leu Met Pro Gly GluArg Asp Glu His 65 70 75 80 Leu Gly His Gly Val Ser Ile Glu Trp Gln TyrLys Lys Tyr Thr Thr 85 90 95 Gln Ile Asp Pro Glu Thr Ala Asp Arg Met IleHis Leu Tyr Tyr Phe 100 105 110 Thr Cys Phe Thr Glu Ser Ala Ile Arg LysAla Ile Leu Gly Gln Arg 115 120 125 Val Leu Thr Lys Cys Glu Tyr Pro AlaGly His Ser Gln Val Gly Thr 130 135 140 Leu Gln Leu Leu Ala Leu Arg ValVal Val Lys Glu Arg Lys His Arg 145 150 155 160 Pro Pro Leu Pro Ser ValGln Lys Leu Thr Glu Asp Arg Trp Asn Lys 165 170 175 His Leu Arg Ile ArgAsp Gln Leu Glu Ser His Ser Met Asn Gly His 180 185 190 <210> SEQ ID NO109 <211> LENGTH: 192 <212> TYPE: PRT <213> ORGANISM: Human <400>SEQUENCE: 109 Met Glu Asn Arg Trp Gln Val Leu Ile Val Trp Gln Val AspArg Gln 1 5 10 15 Lys Val Lys Ala Trp Asn Ser Leu Val Lys Tyr His LysTyr Arg Ser 20 25 30 Arg Lys Thr Gln Asn Trp Asp Tyr Arg His His Tyr GluIle Arg Asn 35 40 45 Pro Arg Ile Ser Ser Gly Val Tyr Ile Pro Val Gly GluAla Lys Ile 50 55 60 Val Val Thr Thr Tyr Trp Gly Leu Met Pro Gly Glu ArgAsp Glu His 65 70 75 80 Leu Gly His Gly Val Ser Ile Glu Trp Gln Tyr LysAsn Tyr Ser Thr 85 90 95 Gln Ile Asp Pro Glu Thr Ala Asp Lys Ile Ile HisLeu His Tyr Phe 100 105 110 Thr Cys Phe Thr Glu Ser Ala Ile Arg Arg AlaIle Leu Gly Gln Arg 115 120 125 Val Leu Thr Arg Cys Glu Tyr Pro Ala GlyHis Ser Gln Val Gly Thr 130 135 140 Leu Gln Leu Leu Ala Leu Arg Ala ValVal Lys Asp Lys Arg Ser Lys 145 150 155 160 Pro Pro Leu Pro Ser Val GlnLys Leu Thr Gly Asp Arg Trp Asn Arg 165 170 175 His Leu Arg Ile Arg AspGln Gln Glu Ser His Ser Met Asn Gly His 180 185 190 <210> SEQ ID NO 110<211> LENGTH: 192 <212> TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE:110 Met Glu Asn Arg Trp Gln Val Met Ile Val Trp Gln Val Asp Arg Met 1 510 15 Arg Ile Arg Thr Trp Asn Ser Leu Val Lys His His Met Tyr Ile Ser 2025 30 Lys Lys Ala Thr Asp Trp Val Tyr Lys His His Tyr Asp Ser Arg His 3540 45 Pro Lys Val Ser Ser Glu Val His Ile Pro Leu Gly Asp Ala Lys Leu 5055 60 Val Ile Arg Thr Tyr Trp Gly Leu His Thr Gly Glu Arg Asp Trp His 6570 75 80 Leu Gly His Gly Val Ser Ile Glu Trp Lys Gln Arg Arg Tyr Ser Thr85 90 95 Gln Ile Asp Pro Asp Leu Ala Asp Gln Leu Ile His Leu His Tyr Phe100 105 110 Asn Cys Phe Ser Glu Ser Ala Ile Arg Lys Ala Ile Leu Gly GlnVal 115 120 125 Val Arg Pro Arg Cys Asp Tyr Pro Ala Gly His Ser Lys ValGly Ser 130 135 140 Leu Gln Tyr Leu Ala Leu Lys Ala Leu Val Thr Pro ThrArg Thr Lys 145 150 155 160 Pro Pro Leu Pro Ser Val Lys Lys Leu Thr GluAsp Arg Trp Asn Lys 165 170 175 Pro Gln Lys Thr Arg Gly His Arg Gly SerGly Pro Met Tyr Gly His 180 185 190 <210> SEQ ID NO 111 <211> LENGTH: 85<212> TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE: 111 Met His HisArg Asp Leu Leu Val Leu Ile Ile Ile Ser Ala Leu Leu 1 5 10 15 Leu IleAsn Val Ile Ile Trp Met Phe Ile Leu Arg Gln Tyr Leu Glu 20 25 30 Gln LysLys Gln Asp Arg Arg Glu Arg Asp Ile Leu Glu Arg Leu Arg 35 40 45 Arg IleAla Glu Ile Lys Asp Asp Ser Asp Tyr Glu Ser Asn Glu Glu 50 55 60 Glu GluGln Glu Val Arg Asp Leu Ile His Ser His Gly Phe Asp Asn 65 70 75 80 ProMet Phe Glu Leu 85 <210> SEQ ID NO 112 <211> LENGTH: 86 <212> TYPE: PRT<213> ORGANISM: Human <400> SEQUENCE: 112 Met Gln Gln Lys Asp Leu LeuLeu Leu Val Ile Ile Ser Ala Leu Leu 1 5 10 15 Leu Ile Asn Ile Ile LeuTrp Met Phe Asn Leu Arg Lys Tyr Leu Glu 20 25 30 Gln Lys Lys Gln Asp ArgArg Glu Arg Glu Ile Leu Glu Arg Ile Arg 35 40 45 Arg Ile Arg Glu Ile ArgAsp Asp Ser Asp Tyr Glu Ser Asn Glu Glu 50 55 60 Glu Glu Gln Glu Val ArgGly His Leu Val His Met Phe Gly Phe Ala 65 70 75 80 Asn Pro Val Phe GluIle 85 <210> SEQ ID NO 113 <211> LENGTH: 85 <212> TYPE: PRT <213>ORGANISM: Human <400> SEQUENCE: 113 Met His His Lys Asp Leu Leu Ile LeuIle Val Ala Ser Ile Leu Leu 1 5 10 15 Phe Thr Asn Ile Val Ile Trp ThrPhe Ile Leu Lys Lys Tyr Leu Glu 20 25 30 Gln Lys Glu Gln Asp Arg Arg GluArg Glu Leu Leu Lys Arg Ile Lys 35 40 45 Arg Ile Arg Glu Val Arg Asp AspSer Asp Tyr Glu Ser Asn Gly Asp 50 55 60 Gly Gly Gln Glu Val Ile His LeuVal His Thr His Gly Phe Val Asn 65 70 75 80 Pro Met Phe Glu Leu 85 <210>SEQ ID NO 114 <211> LENGTH: 85 <212> TYPE: PRT <213> ORGANISM: Human<400> SEQUENCE: 114 Met Gln Leu Arg Asp Gln Leu Thr Leu Ile Ile Ile SerAla Leu Leu 1 5 10 15 Leu Val Asn Val Val Leu Trp Thr Phe Ile Leu ArgGln Tyr Leu Lys 20 25 30 Gln Lys Lys Gln Asp Arg Arg Gly Arg Glu Ile LeuGlu Arg Leu Arg 35 40 45 Arg Ile Arg Gln Ile Glu Asp Asp Ser Asp Tyr GluSer Asp Gly Lys 50 55 60 Glu Glu Gln Glu Val Arg Asp Leu Val His Ser TyrGly Phe Asp Asn 65 70 75 80 Pro Met Phe Glu Pro 85 <210> SEQ ID NO 115<211> LENGTH: 100 <212> TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE:115 Met Glu Arg Ala Pro Glu Asp Gln Gly Pro Ala Arg Glu Pro Phe Asn 1 510 15 Glu Trp Ala Leu Glu Ile Leu Glu Glu Leu Lys Ala Glu Ala Val Arg 2025 30 His Phe Pro Arg Gln Trp Leu Gln Ala Leu Gly Gln Tyr Ile Tyr Glu 3540 45 Thr Tyr Gly Asp Thr Trp Val Gly Val Met Ala Ile Thr Arg Ile Leu 5055 60 Gln Gln Ile Leu Phe Ala His Phe Arg Ile Gly Cys Gln His Ser Arg 6570 75 80 Ile Gly Ile Asn Pro Thr Asn Thr Arg Gly Arg Gly Arg Arg Asn Gly85 90 95 Ser Ser Arg Ser 100 <210> SEQ ID NO 116 <211> LENGTH: 100 <212>TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE: 116 Met Glu Gln Ala ProGlu Asp Gln Gly Pro Ala Arg Glu Pro Phe Asn 1 5 10 15 Glu Trp Thr LeuGlu Leu Leu Glu Glu Leu Lys Ala Glu Ala Val Arg 20 25 30 His Phe Pro ArgPro Trp Leu Gln Ala Leu Gly Gln Tyr Ile Tyr Asp 35 40 45 Thr Tyr Gly AspThr Trp Val Gly Val Met Ala Ile Ile Arg Leu Leu 50 55 60 Gln Leu Met IlePhe Ala His Phe Arg Ile Gly Cys Gln His Ser Arg 65 70 75 80 Ile Gly IleAsn Pro Ser Asn Thr Arg Gly Arg Gly Arg Arg Asn Gly 85 90 95 Ser Ser ArgPro 100 <210> SEQ ID NO 117 <211> LENGTH: 93 <212> TYPE: PRT <213>ORGANISM: Human <400> SEQUENCE: 117 Met Glu Gln Ala Pro Glu Asn Gln GlyPro Ala Arg Glu Pro Phe Asn 1 5 10 15 Glu Trp Ala Leu Glu Thr Leu GluGlu Ile Lys Ala Glu Ala Val Arg 20 25 30 His Phe Pro Arg Pro Trp Leu GlnSer Leu Gly Gln Tyr Ile Tyr Glu 35 40 45 Thr Tyr Gly Asp Thr Trp Glu GlyVal Met Ala Ile Ile Arg Ile Leu 50 55 60 Gln Gln Leu Ile Phe Ala His PheArg Ile Gly Cys Gln His Ser Arg 65 70 75 80 Ile Gly Ile Thr Pro Ser AsnAla Arg Gly Arg Gly Arg 85 90 <210> SEQ ID NO 118 <211> LENGTH: 100<212> TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE: 118 Met Glu GlnAla Pro Glu Asp Gln Gly Ala Gln Arg Glu Arg Ser Asn 1 5 10 15 Val TrpThr Leu Asp Leu Leu Glu Glu Leu Lys His Glu Ala Val Arg 20 25 30 His PhePro Arg Pro Trp Leu Gln Gly Leu Gly Gln Tyr Ile Tyr Glu 35 40 45 Thr TyrGly Asp Thr Trp Glu Gly Val Glu Ala Ile Ile Arg Ile Leu 50 55 60 Gln GlnLeu Leu Phe Ala His Phe Arg Ile Gly Cys Gln His Ser Arg 65 70 75 80 IleGly Ile Asn Pro Ser Asn Pro Arg Gly Lys Gly Arg Arg Asn Gly 85 90 95 SerSer Arg Ser 100 <210> SEQ ID NO 119 <211> LENGTH: 71 <212> TYPE: PRT<213> ORGANISM: Human <400> SEQUENCE: 119 Met Asp Pro Val Asp Pro GluMet Pro Pro Trp His His Pro Gly Ser 1 5 10 15 Gln Pro Gln Ile Pro CysAsn Asn Cys Tyr Cys Lys Arg Cys Cys Tyr 20 25 30 His Cys Leu Val Cys PheThr Arg Lys Gly Leu Gly Ile Ser Tyr Gly 35 40 45 Arg Lys Lys Arg Arg GlnArg Arg Ala Ala Ala Ser His Pro Asp Asn 50 55 60 Lys Asp Leu Val Pro GluGln 65 70 <210> SEQ ID NO 120 <211> LENGTH: 71 <212> TYPE: PRT <213>ORGANISM: Human <400> SEQUENCE: 120 Met Asp Pro Val Asp Pro Glu Met ProPro Trp His His Pro Gly Ser 1 5 10 15 Gln Pro Gln Asn Pro Cys Asn LysCys Tyr Cys Lys Lys Cys Cys Tyr 20 25 30 His Cys Tyr Val Cys Phe Thr SerLys Gly Leu Gly Ile Ser Tyr Gly 35 40 45 Arg Lys Lys Arg Arg Arg Pro AlaAla Ala Ala Ser Arg Pro Asp Asn 50 55 60 Lys Asp Leu Val Pro Glu Gln 6570 <210> SEQ ID NO 121 <211> LENGTH: 71 <212> TYPE: PRT <213> ORGANISM:Human <400> SEQUENCE: 121 Met Asp Pro Val Asp Pro Glu Val Pro Pro TrpHis His Pro Gly Ser 1 5 10 15 Gln Pro Pro Thr Pro Cys Asn Ala Cys TyrCys Lys Arg Cys Cys Tyr 20 25 30 His Cys Tyr Leu Cys Phe Thr Lys Lys GlyLeu Gly Ile Ser His Gly 35 40 45 Arg Lys Lys Arg Arg Arg Pro Ala Ala AlaAla Ser Ser Ser Asn Asn 50 55 60 Lys Asp Leu Val Pro Glu Gln 65 70 <210>SEQ ID NO 122 <211> LENGTH: 71 <212> TYPE: PRT <213> ORGANISM: Human<400> SEQUENCE: 122 Met Asp Pro Val Asp Pro Glu Ile Pro Pro Trp His HisPro Gly Ser 1 5 10 15 Gln Pro Gln Thr Pro Cys Asn Asn Cys Ser Cys LysLys Cys Cys Tyr 20 25 30 His Cys Tyr Val Cys Phe Thr Arg Lys Gly Leu GluIle Ser Tyr Gly 35 40 45 Arg Lys Lys Arg Arg Arg Ser Ala Ala Glu Thr ArgHis Pro Asp Asn 50 55 60 Gln Asp Ile Val Pro Glu Gln 65 70 <210> SEQ IDNO 123 <211> LENGTH: 13 <212> TYPE: PRT <213> ORGANISM: Human <400>SEQUENCE: 123 Ala Ile Arg Ile Ile Lys Ile Leu Tyr Gln Ser Ser Lys 1 5 10<210> SEQ ID NO 124 <211> LENGTH: 26 <212> TYPE: PRT <213> ORGANISM:Human <400> SEQUENCE: 124 Met Ala Gly Arg Ser Asp Asp Asp Gln Gln LeuLeu Gln Ala Ala Arg 1 5 10 15 Ile Ile Lys Ile Leu Tyr Gln Ser Ser Lys 2025 <210> SEQ ID NO 125 <211> LENGTH: 26 <212> TYPE: PRT <213> ORGANISM:Human <400> SEQUENCE: 125 Met Ala Gly Arg Ser Asp Asp Asp Gln Gln LeuLeu Gln Ala Leu Arg 1 5 10 15 Ile Ile Lys Ile Leu Tyr Gln Ser Ser Lys 2025 <210> SEQ ID NO 126 <211> LENGTH: 26 <212> TYPE: PRT <213> ORGANISM:Human <400> SEQUENCE: 126 Met Ala Gly Arg Ser Asp Glu Asp Gln Pro LeuLys Arg Val Ile Gln 1 5 10 15 Ile Ile Lys Ile Leu Tyr Gln Ser Ser Lys 2025 <210> SEQ ID NO 127 <211> LENGTH: 42 <212> TYPE: PRT <213> ORGANISM:Human <400> SEQUENCE: 127 Ile Lys Val Val Pro Arg Arg Lys Ala Lys IleIle Arg His Tyr Gly 1 5 10 15 Lys Gln Met Ala Gly Ala Asp Ser Met AlaSer Gly Gln Thr Glu Ser 20 25 30 Glu Ser Val Glu Gln Pro Gly Glu Ile Pro35 40 <210> SEQ ID NO 128 <211> LENGTH: 42 <212> TYPE: PRT <213>ORGANISM: Human <400> SEQUENCE: 128 Ile Lys Val Val Pro Arg Arg Lys AlaLys Ile Ile Arg Asp Tyr Gly 1 5 10 15 Lys Gln Met Ala Gly Thr Asp SerMet Ala Ser Arg Gln Thr Glu Ser 20 25 30 Glu Asn Val Glu Gln Leu Gly GluIle Pro 35 40 <210> SEQ ID NO 129 <211> LENGTH: 36 <212> TYPE: PRT <213>ORGANISM: Human <400> SEQUENCE: 129 Ile Lys Val Val Pro Arg Arg Lys AlaLys Ile Leu Arg Asp Tyr Gly 1 5 10 15 Lys Gln Met Ala Gly Ala Asp SerMet Ala Ser Gly Gln Thr Glu Ser 20 25 30 Glu Ser Met Glu 35 <210> SEQ IDNO 130 <211> LENGTH: 106 <212> TYPE: PRT <213> ORGANISM: Human <400>SEQUENCE: 130 His Asn Phe Lys Arg Lys Gly Gly Ile Gly Gly Tyr Ser AlaGly Glu 1 5 10 15 Arg Ile Ile Asp Ile Ile Ala Ser Asp Ile Gln Thr LysGlu Leu Gln 20 25 30 Lys Gln Ile Thr Lys Ile Gln Asn Phe Arg Val Tyr TyrArg Asp Ser 35 40 45 Arg Asp Pro Ile Trp Lys Gly Pro Ala Lys Leu Leu TrpLys Gly Glu 50 55 60 Gly Ala Val Val Ile Gln Asp Asn Ser Asp Ile Lys ValVal Pro Arg 65 70 75 80 Arg Lys Ala Lys Ile Ile Lys Asp Tyr Gly Lys GlnMet Ala Gly Asp 85 90 95 Asp Cys Val Ala Ser Arg Gln Asn Glu Asp 100 105<210> SEQ ID NO 131 <211> LENGTH: 35 <212> TYPE: PRT <213> ORGANISM:Human <400> SEQUENCE: 131 Met Ile Val Thr Met Lys Ala Met Lys Arg ArgAsn Arg Lys Leu Glu 1 5 10 15 Ile Leu Tyr Ile Val Met Ala Leu Ile IlePro Cys Leu Ser Ser Asp 20 25 30 Gln Lys Tyr 35 <210> SEQ ID NO 132<211> LENGTH: 36 <212> TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE:132 Met Ile Val Thr Met Lys Ala Met Lys Arg Lys Asn Lys Lys Leu Gly 1 510 15 Val Ile Leu Cys Ile Cys Leu Ala Leu Leu Ile Pro Cys Leu Arg Ser 2025 30 Asn Asp Leu Tyr 35 <210> SEQ ID NO 133 <211> LENGTH: 35 <212>TYPE: PRT <213> ORGANISM: Human <400> SEQUENCE: 133 Met Ile Val Ile MetLys Ala Met Glu Met Glu Asp Lys Lys Leu Tyr 1 5 10 15 Ile Leu Cys IleLeu Met Val Leu Leu Thr Pro Cys Leu Ser Ser Asp 20 25 30 Lys Leu Tyr 35<210> SEQ ID NO 134 <211> LENGTH: 715 <212> TYPE: PRT <213> ORGANISM:Human <400> SEQUENCE: 134 Met Thr Val Thr Met Lys Ala Met Glu Lys ArgAsn Arg Lys Leu Gly 1 5 10 15 Ile Leu Cys Ile Val Met Ala Leu Ile ThrPro Cys Leu Ser His Asp 20 25 30 Gln Arg Tyr Ala Thr Val Tyr Ala Gly ValPro Val Trp Glu Glu Ala 35 40 45 Asn Pro Val Leu Phe Cys Ala Ser Asp AlaAsn Leu Thr Ser Thr Glu 50 55 60 Lys His Asn Ile Trp Ala Ser Gln Ala CysVal Pro Thr Asp Pro Thr 65 70 75 80 Pro His Glu Tyr Pro Leu His Asn ValThr Asp Asn Phe Asn Ile Trp 85 90 95 Lys Asn Tyr Met Val Glu Gln Met GlnAsp Asp Ile Ile Ser Leu Trp 100 105 110 Glu Gln Ser Leu Lys Pro Cys ValGln Met Thr Phe Leu Cys Val Gln 115 120 125 Met Asn Cys Thr Ser Val SerAsn Ser Ser Val Ser Asn Ser Ser Val 130 135 140 Ser Asn Ser Ser Val SerAsn Ser Ser Val Ser Asp Ser Thr Ile Pro 145 150 155 160 Lys Lys Lys AsnAsn Ser Ser Ser Glu Asp Leu Leu Lys Gln Cys Asp 165 170 175 Phe Asn AlaThr Thr Val Leu Lys Asp Lys Lys Glu Lys Lys Gln Thr 180 185 190 Leu PheTyr Val Ser Asp Leu Met Lys Leu Thr Asn Val Thr Asn Asp 195 200 205 ThrMet Tyr Thr Leu Ile Asn Cys Asn Ser Thr Thr Ile Lys Gln Ala 210 215 220Cys Pro Lys Val Thr Phe Glu Pro Ile Pro Ile His Tyr Cys Ala Pro 225 230235 240 Ala Gly Tyr Ala Ile Phe Lys Cys Asn Asn Thr Glu Phe Asn Gly Thr245 250 255 Gly Pro Cys Asn Asn Ile Thr Val Val Thr Cys Thr His Gly IleArg 260 265 270 Pro Thr Val Ser Thr Gln Leu Ile Leu Asn Gly Thr Leu SerGlu Gly 275 280 285 Lys Ile Arg Ile Met Gly Arg Asn Ile Thr Asp Ser GlyLys Asn Ile 290 295 300 Ile Val Thr Leu Asn Tyr Thr Ile Asn Ile Thr CysGlu Arg Thr Trp 305 310 315 320 Asn Gln Ser Val Gln Glu Ile Pro Ile GlyPro Met Ala Trp Tyr Ser 325 330 335 Met Ser Val Glu Lys Asp Lys Asn ThrThr Gly Ser Arg Ser Ala Asp 340 345 350 Cys Gln Tyr Asn Thr Ser Glu TrpThr Arg Ala Leu Glu Gln Thr Ala 355 360 365 Glu Arg Tyr Leu Glu Leu MetAsn Asn Thr Gly Asn Thr Asp Asn Thr 370 375 380 Thr Val Ile Phe Asn HisSer Thr Gly Gly Asp Pro Glu Val Ser Phe 385 390 395 400 Leu His Phe AsnCys His Gly Glu Phe Phe Tyr Cys Asn Thr Ser Gly 405 410 415 Met Phe AsnTyr Thr Phe Ser Cys Lys Gly Thr Asn Cys Thr Gln Val 420 425 430 Gly SerGln Asn Glu Tyr Asn Asn His Thr Thr Lys Ile Pro Cys Arg 435 440 445 IleLys Gln Val Val Arg Ser Trp Ile Arg Gly Gly Ser Gly Leu Tyr 450 455 460Ala Pro Pro Arg Gln Gly Pro Leu Lys Cys Ser Ser Asn Ile Thr Gly 465 470475 480 Met Ile Leu Gln Leu Asp Lys Pro Trp Asn Arg Ser Gly His Asn Asn485 490 495 Asp Thr Thr Phe Arg Pro Ile Gly Gly Glu Met Lys Asp Ile TrpArg 500 505 510 Thr Glu Leu Phe Lys Tyr Lys Val Val Lys Val Lys Pro PheSer Val 515 520 525 Ala Pro Thr Lys Ile Ala Arg Pro Val Ile Gly Thr GlyThr Gln Arg 530 535 540 Glu Lys Arg Ala Val Gly Leu Gly Met Leu Phe LeuGly Val Leu Ser 545 550 555 560 Ala Ala Gly Ser Thr Met Gly Ala Ala AlaThr Thr Leu Ala Val Gln 565 570 575 Thr His Thr Leu Met Lys Gly Ile ValGln Gln Gln Asp Asn Leu Leu 580 585 590 Arg Ala Ile Gln Ala Gln Gln GlnLeu Leu Arg Leu Ser Val Trp Gly 595 600 605 Ile Arg Gln Leu Arg Ala ArgLeu Leu Ala Leu Glu Thr Leu Ile Gln 610 615 620 Asn Gln Gln Leu Leu AsnLeu Trp Gly Cys Lys Gly Arg Leu Val Cys 625 630 635 640 Tyr Thr Ser ValGln Trp Asn Arg Thr Trp Thr Asn Asn Thr Asn Leu 645 650 655 Asp Ser IleTrp Glu Asn Leu Thr Trp Gln Glu Trp Asp Gln Gln Ile 660 665 670 Ser AsnIle Ser Ser Thr Ile Tyr Glu Glu Ile Gln Lys Ala Gln Ile 675 680 685 GlnGln Glu Tyr Asn Glu Lys Lys Leu Leu Glu Leu Asp Glu Trp Ala 690 695 700Ser Ile Trp Asn Trp Leu Asp Ile Thr Lys Cys 705 710 715 <210> SEQ ID NO135 <211> LENGTH: 38 <212> TYPE: PRT <213> ORGANISM: Human <220>FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(38) <400>SEQUENCE: 135 Cys Glu Arg Thr Trp Asn Gln Ser Val Gln Glu Ile Pro IleGly Pro 1 5 10 15 Met Ala Trp Tyr Ser Met Ser Val Glu Leu Asp Leu AsnThr Thr Gly 20 25 30 Ser Arg Ser Ala Asp Cys 35 <210> SEQ ID NO 136<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Human <220> FEATURE:<221> NAME/KEY: PEPTIDE <222> LOCATION: (1)..(7) <400> SEQUENCE: 136 SerVal Gln Glu Ile Pro Ile 1 5 <210> SEQ ID NO 137 <211> LENGTH: 23 <212>TYPE: PRT <213> ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE<222> LOCATION: (1)..(23) <400> SEQUENCE: 137 Asn Gln Gln Leu Leu AsnLeu Trp Gly Cys Lys Gly Arg Leu Val Cys 1 5 10 15 Tyr Thr Ser Val GlnTrp Asn 20 <210> SEQ ID NO 138 <211> LENGTH: 24 <212> TYPE: PRT <213>ORGANISM: Human <220> FEATURE: <221> NAME/KEY: PEPTIDE <222> LOCATION:(1)..(24) <400> SEQUENCE: 138 Asn Gln Gln Leu Leu Asn Leu Trp Gly CysLys Gly Arg Leu Val Cys 1 5 10 15 Tyr Thr Ser Val Lys Trp Asn Asn 20<210> SEQ ID NO 139 <211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM:Human <400> SEQUENCE: 139 gggttcttgg gagcagcagg aagcactatg ggcg 34 <210>SEQ ID NO 140 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Human<400> SEQUENCE: 140 tctgaaacga cagaggtgag tatccctgcc taa 33 <210> SEQ IDNO 141 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Human <400>SEQUENCE: 141 tggatcccac agtgtactga agggtatagt gca 33 <210> SEQ ID NO142 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Human <400>SEQUENCE: 142 catttagtta tgtcaagcca attccaaa 28 <210> SEQ ID NO 143<211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE:143 gttctccata tatctttcat atctccccct a 31 <210> SEQ ID NO 144 <211>LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 144ttgtacacat ggcattaggc caacagtaag t 31 <210> SEQ ID NO 145 <211> LENGTH:31 <212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 145 tgaattcctaatattgaatg ggacactctc t 31 <210> SEQ ID NO 146 <211> LENGTH: 32 <212>TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 146 tggatcctacaataaaagaa ttctccatga ca 32 <210> SEQ ID NO 147 <211> LENGTH: 24 <212>TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 147 gggtttattacagggacagc agag 24 <210> SEQ ID NO 148 <211> LENGTH: 24 <212> TYPE: DNA<213> ORGANISM: Human <400> SEQUENCE: 148 ggttggggtc tgtgggtaca cagg 24<210> SEQ ID NO 149 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM:Human <400> SEQUENCE: 149 gcaaaactac tctggaaagg tg 22 <210> SEQ ID NO150 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Human <400>SEQUENCE: 150 gcwtctttcc acacaggtac ccc 23 <210> SEQ ID NO 151 <211>LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 151catattgggg attgatgcca g 21 <210> SEQ ID NO 152 <211> LENGTH: 20 <212>TYPE: DNA <213> ORGANISM: Human <400> SEQUENCE: 152 gcatyagcgttacttactgc 20

What is claimed is:
 1. An isolated antigen from the HIV-1 group O straingp160 env precursor protein comprising the amino acid sequence of SEQ IDNO:100.
 2. A method for detecting anti-HIV-1 antibodies in a samplecomprising: a) contacting the sample with an isolated antigen from theHIV-1 group O strain gp160 env precursor protein comprising the aminoacid sequence of SEQ ID NO:100, b) allowing the isolated antigen andanti-HIV antibodies to interact, and c) detecting the interactionbetween the antigen and the anti-HIV antibodies.
 3. A kit for detectingHIV-1 antibodies comprising an isolated antigen from the HIV-1 group Ostrain gp160 env precursor protein comprising the amino acid sequence ofSEQ ID NO:100.
 4. An immunogenic composition comprising: a) an isolatedantigen from the HIV-1 group O strain gp160 env precursor protein whichcomprises the amino acid sequence of SEQ ID NO:100; and b) apharmaceutically acceptable carrier.